Dipeptidomimetics as inhibitors of human immunoproteasomes

ABSTRACT

The compounds of the present invention are represented by the following compounds having Formula I: 
     
       
         
         
             
             
         
       
     
     where the substituents R 1 , R 4 , L, M, X, Y, and s are as defined herein. The compounds of the present invention are also represented by the following compounds having Formula (Ia), Formula (Ib), or Formula (Ic): 
     
       
         
         
             
             
         
       
     
     where the substituents R 1 -R 4 , R x , R y , X, Y, and s are as defined herein. 
     These compounds are used in the treatment of cancer, immunologic disorders, autoimmune disorders, neurodegenerative disorders, or inflammatory disorders or for providing immunosuppression for transplanted organs or tissues.

This application claims the benefit of U.S. Provisional PatentApplication Ser. No. 62/038,636, filed Aug. 18, 2014, which is herebyincorporated by reference in its entirety.

This invention was made with government support under grant number5R21AI101393 awarded by the National Institutes of Health. Thegovernment has certain rights in this invention.

FIELD OF THE INVENTION

The present invention relates to inhibitors of human immunoproteasomes.

BACKGROUND OF THE INVENTION

The proteasome is a large, ATP-dependent, multi-subunit, barrel-shapedN-terminal nucleophile hydrolase present in the cytosol and nucleus ofeukaryotic cells, and is responsible for the degradation of the majorityof cellular proteins (Baumeister et al., “The Proteasome: Paradigm of aSelf-Compartmentalizing Protease,” Cell 92:367-380 (1998); Goldberg, A.L., “Functions of the Proteasome: From Protein Degradation and ImmuneSurveillance to Cancer Therapy,” Biochemical Society Transactions35:12-17 (2007)). The proteasome not only controls many criticalcellular checkpoints through degradation, but also generates peptidesfor antigen presentation (Goldberg, A. L., “Functions of the Proteasome:From Protein Degradation and Immune Surveillance to Cancer Therapy,”Biochemical Society Transactions 35:12-17 (2007); Rock et al.,“Inhibitors of the Proteasome Block the Degradation of Most CellProteins and the Generation of Peptides Presented on MHC Class IMolecules,” Cell 78:761-771 (1994)). Highly specific proteasomeinhibitors can markedly limit the overall supply of peptides for MHCclass I molecules and thus block antigen presentation (Rock et al.,“Protein Degradation and the Generation of MHC Class I-PresentedPeptides,” Advances in Immunology 80:1-70 (2002)). The constitutiveproteasome core particle is called 20S (c-20S) because of itssedimentation properties. Inside the c-20S core reside two copies ofeach of three proteases with distinct specificities, β1 (caspase-like),β2 (tryptic-like) and β5 (chymotryptic-like) (Bedford et al.,“Ubiquitin-Like Protein Conjugation and the Ubiquitin-Proteasome Systemas Drug Targets,” Nature Reviews. Drug Discovery 10:29-46 (2011)).However, lymphocytes and cells that have responded to interferon-γexpress a different proteasome, called the immunoproteasome (i-20S), inwhich the corresponding proteases are the products of different genes:β1i, β2i and β5i. Intermediate proteasomes that contain mixed β subunitsare found in many cells, for example in the mucosa of the colon andsmall bowel (Guillaume et al., “Two Abundant Proteasome Subtypes thatUniquely Process Some Antigens Presented by HLA Class I Molecules,”Proc. Nat'l Acad. Sci. USA 107:18599-18604 (2010)). The effects ofreplacement of constitutive subunits by immuno-β subunits includeincreased proteolytic activity and altered peptide preferences of theactive sites (Rock et al., “Proteases in MHC Class I Presentation andCross-Presentation,” Journal of Immunology 184:9-15d (2010)). Forexample, the caspase-like β1 replacement, β1i, preferentially cleavesafter small hydrophobic residues rather than after aspartate (Huber etal., “Immuno- and Constitutive Proteasome Crystal Structures RevealDifferences in Substrate and Inhibitor Specificity,” Cell 148:727-738(2012)). This results in altered peptide products, such that mice withcombined deficiency of β1i, β2i, and β5i are viable, fertile and healthybut express a different antigenic peptide repertoire than wild typemice, as evidenced by their rejection of syngeneic wild type splenocytes(Kincaid et al., “Mice Completely Lacking Immunoproteasomes Show MajorChanges in Antigen Presentation,” Nature Immunology 13:129-135 (2012)).Hu c-20S and i-20S appear to regulate cytokine production throughdifferent pathways (Muchamuel et al., “A Selective Inhibitor of theImmunoproteasome Subunit LMP7 Blocks Cytokine Production and AttenuatesProgression of Experimental Arthritis,” Nature Medicine 15:781-787(2009)). Hu c-20S controls the activation of NF-κB via the degradationof IκB, the binding partner of NF-κB in the cytosol (Perkins, N. D.,“Integrating Cell-Signalling Pathways with NF-[Kappa]B and IKKFunction,” Nat. Rev. Mol. Cell Biol. 8:49-62 (2007)), and inhibition ofc-20S blocks the activation of NF-κB (Meng et al., “Epoxomicin, a Potentand Selective Proteasome Inhibitor, Exhibits In Vivo AntiinflammatoryActivity,” Proc. Nat'l Acad. Sci. USA 96:10403-10408 (1999)). For itspart, among other potential pathways, i-20S appears to control theco-translocation of TLR9 and Unc93B1, an endoplasmic reticulum(ER)-resident protein, to endosomes (Hirai et al., “BortezomibSuppresses Function and Survival of Plasmacytoid Dendritic Cells byTargeting Intracellular Trafficking of Toll-Like Receptors andEndoplasmic Reticulum Homeostasis,” Blood 117:500-509 (2011)).Proteasomes control diverse cellular functions, among them signaltransduction for inflammatory cytokine release, antigen presentation,and the ability of plasma cells to secrete antibodies without dying fromaccumulation of misfolded immunoglobulins (Goldberg, A. L., “Functionsof the Proteasome: From Protein Degradation and Immune Surveillance toCancer Therapy,” Biochemical Society Transactions 35:12-17 (2007);Bedford et al., “Ubiquitin-Like Protein Conjugation and theUbiquitin-Proteasome System as Drug Targets,” Nature Reviews. DrugDiscovery 10:29-46 (2011); Neubert et al., “The Proteasome InhibitorBortezomib Depletes Plasma Cells and Protects Mice with Lupus-LikeDisease from Nephritis,” Nature Medicine 14:748-755 (2008)). Thus theproteasome could be a target for treating autoimmune and inflammatorydiseases. For example, inhibition of the proteasome in plasmacytoiddendritic cells (pDCs) prevents the trafficking of TLRs, resulting in ablock of nuclear translocation of IRF-7, consequently suppressing theproduction of IFNα (Hirai et al., “Bortezomib Suppresses Function andSurvival of Plasmacytoid Dendritic Cells by Targeting IntracellularTrafficking of Toll-Like Receptors and Endoplasmic ReticulumHomeostasis,” Blood 117:500-509 (2011)), a cytokine implicated insystemic lupus erythematosus (SLE). However, by the same token,widespread inhibition of proteasomes can be expected to be toxic and hasproven toxic in the clinic.

Two proteasome inhibitors approved by the FDA for treatment ofmalignancy, Bortezomib and Carfilzomib, inhibit both the c-20S β5c andthe i-20S β5i (Huber et al., “Inhibitors for the Immuno- andConstitutive Proteasome: Current and Future Trends in Drug Development,”Angewandte Chemie 51:8708-8720 (2012)). Bortezomib, a dipeptidylboronate, is a slow-binding, covalent but reversible inhibitor, whereasCarfilzomib is a peptide with an epoxyketone warhead that inhibitsproteasomes irreversibly. In addition to treatment of malignancy,Bortezomib has been reported to be effective in inflammatory boweldisease (IBD), SLE, graft-versus-host disease, antibody-mediated graftrejection, rheumatoid arthritis (RA), and other immunologic, autoimmuneand/or inflammatory conditions. However, such a broad-spectrum inhibitoris too toxic for chronic treatment of non-malignant diseases. ONX 0914,another peptide epoxyketone, has modest selectivity for i-20S β5i(Muchamuel et al., “A Selective Inhibitor of the ImmunoproteasomeSubunit LMP7 Blocks Cytokine Production and Attenuates Progression ofExperimental Arthritis,” Nature Medicine 15:781-787 (2009)) and isreported to have efficacy in rheumatoid arthritis (Muchamuel et al., “ASelective Inhibitor of the Immunoproteasome Subunit LMP7 Blocks CytokineProduction and Attenuates Progression of Experimental Arthritis,” NatureMedicine 15:781-787 (2009)), SLE (Ichikawa et al., “Beneficial Effect ofNovel Proteasome Inhibitors in Murine Lupus Via Dual Inhibition of TypeI Interferon and Autoantibody-Secreting Cells,” Arthritis and Rheumatism64:493-503 (2012)), experimental colitis (Basler et al., “Prevention ofExperimental Colitis by a Selective Inhibitor of the Immunoproteasome,”Journal of Immunology 185:634-641 (2010)), and multiple sclerosis(Basler et al., “Inhibition of the immunoproteasome amelioratesexperimental autoimmune encephalomyelitis,” EMBO Mol. Med. 6:226-238(2014)). Nonetheless, it, too, acts irreversibly and has considerabletoxicity.

The present invention is directed to overcoming these and otherdeficiencies in the art.

SUMMARY OF THE INVENTION

One aspect of the present invention relates to a compound of Formula(I):

wherein

L is —(CR³R^(x))_(p)—;

M is —(CR²R^(y))_(r)—;

R¹ is selected from the group consisting of monocyclic and bicyclicaryl, biphenyl, monocyclic and bicyclic heteroaryl, monocyclic andbicyclic heterocyclyl, and monocyclic and bicyclic non-aromaticheterocycle, wherein monocyclic and bicyclic aryl, biphenyl, monocyclicand bicyclic heteroaryl, monocyclic and bicyclic heterocyclyl, andmonocyclic and bicyclic non-aromatic heterocycle can be optionallysubstituted from 1 to 3 times with a substituent selected independentlyat each occurrence thereof from the group consisting of halogen, cyano,—CF₃, C₁₋₆ alkyl, and C₁₋₆ alkoxy;

R² is independently selected at each occurrence thereof from the groupconsisting of H, D, C₁₋₆ alkyl, —CH₂OC₁₋₆ alkyl, —CH₂Ar, and—CH₂heteroaryl, wherein aryl (Ar) can be optionally substituted from 1to 3 times with a substituent selected independently at each occurrencethereof from the group consisting of halogen, cyano, C₁₋₆ alkyl, andC₁₋₆ alkoxy;

R³ is independently selected at each occurrence thereof from the groupconsisting of H, D, —CH₂OC₁₋₆ alkyl, —(CH₂)_(m)C(O)NHR⁵,—(CH₂)_(m)C(O)NR⁶R⁷, —(CH₂)_(m)C(O)OH, and —(CH₂)_(m)C(O)OBn;

R⁴ is selected from the group consisting of H, —C(O)(CH₂)_(n)Ph,—C(O)CH₂NR⁶R⁷, —SO₂Ar, —SO₂C₁₋₆ alkyl, —SO₂C₃₋₆ cycloalkyl,—C(O)(CH₂)_(n)Het, —C(O)C(O)Het, —C(O)C₁₋₆ alkyl, —C(O)OC₁₋₆ alkyl,—C(O)CF₃, heteroaryl, and —(CH₂)_(n)NR⁶R⁷, wherein aryl (Ar) andheteroaryl (Het) can be optionally substituted from 1 to 3 times with asubstituent selected independently at each occurrence thereof from thegroup consisting of halogen, cyano, C₁₋₆ alkyl, and C₁₋₆ alkoxy;

R⁵ is selected from the group consisting of C₁₋₆ alkyl, C₁₋₆ alkoxy,non-aromatic heterocycle, —NR⁶R⁷, and —CR⁸R⁹;

R⁶, R⁷, R⁸, and R⁹ are each independently selected from the groupconsisting of H, D, C₁₋₆ alkyl, and —(CH₂)_(k)OH;

or R⁶ and R⁷ are taken together with the nitrogen to which they areattached to form a morpholine ring;

or R⁸ and R⁹ are taken together with the carbon to which they areattached to form an oxetane ring;

R^(x) is independently selected at each occurrence thereof from thegroup consisting of H, D, —CH₂OC₁₋₆ alkyl, —(CH₂)_(m)C(O)NHR⁵,—(CH₂)_(m)C(O)NR⁶R⁷, and —CH₂C(O)R⁵ _(;)

R^(y) is independently selected at each occurrence thereof from thegroup consisting of H, D, C₁₋₆ alkyl, —CH₂OC₁₋₆ alkyl, —CH₂Ar, and—CH₂heteroaryl, wherein aryl (Ar) can be optionally substituted from 1to 3 times with a substituent selected independently at each occurrencethereof from the group consisting of halogen, cyano, C₁₋₆ alkyl, andC₁₋₆ alkoxy;

X is —(CH₂)_(q)—, —O—, or —(CD₂)_(q)—;

Y is O or S;

k is 1, 2, or 3;

m is 0, 1, 2, 3, 4, or 5;

n is 0, 1, 2, or 3;

p is 1 or 2;

q is 0, 1, or 2;

r is 1 or 2; and

s is 0 or 1;

with a proviso that when s is 0, then r is 2; and when s is 1, thenr+p≥3,

or an oxide thereof, a pharmaceutically acceptable salt thereof, asolvate thereof, or a prodrug thereof.

Another aspect of the present invention relates to a method of treatingcancer, immunologic disorders, autoimmune disorders, neurodegenerativedisorders, or inflammatory disorders in a subject or for providingimmunosuppression for transplanted organs or tissues in a subject. Thismethod includes administering to the subject in need thereof a compoundof the Formula (I):

wherein

L is —(CR³R^(x))_(p)—;

M is —(CR²R^(y))_(r)—;

R¹ is selected from the group consisting of monocyclic and bicyclicaryl, biphenyl, monocyclic and bicyclic heteroaryl, monocyclic andbicyclic heterocyclyl, and monocyclic and bicyclic non-aromaticheterocycle, wherein monocyclic and bicyclic aryl, biphenyl, monocyclicand bicyclic heteroaryl, monocyclic and bicyclic heterocyclyl, andmonocyclic and bicyclic non-aromatic heterocycle can be optionallysubstituted from 1 to 3 times with a substituent selected independentlyat each occurrence thereof from the group consisting of halogen, cyano,—CF₃, C₁₋₆ alkyl, and C₁₋₆ alkoxy;

R² is independently selected at each occurrence thereof from the groupconsisting of H, D, C₁₋₆ alkyl, —CH₂OC₁₋₆ alkyl, —CH₂Ar, and—CH₂heteroaryl, wherein aryl (Ar) can be optionally substituted from 1to 3 times with a substituent selected independently at each occurrencethereof from the group consisting of halogen, cyano, C₁₋₆ alkyl, andC₁₋₆ alkoxy;

R³ is independently selected at each occurrence thereof from the groupconsisting of H, D, —CH₂OC₁₋₆ alkyl, —(CH₂)_(m)C(O)NHR⁵,—(CH₂)_(m)C(O)NR⁶R⁷, —(CH₂)_(m)C(O)OH, and —(CH₂)_(m)C(O)OBn_(;)

R⁴ is selected from the group consisting of H, —C(O)(CH₂)_(n)Ph,—C(O)CH₂NR⁶R⁷, —SO₂Ar, SO₂C₁₋₆ alkyl, —SO₂C₃₋₆ cycloalkyl,—C(O)(CH₂)_(n)Het, —C(O)C(O)Het, —C(O)C₁₋₆ alkyl, —C(O)OC₁₋₆ alkyl,—C(O)CF₃, heteroaryl, and —(CH₂)_(n)NR⁶R⁷, wherein aryl (Ar) andheteroaryl (Het) can be optionally substituted from 1 to 3 times with asubstituent selected independently at each occurrence thereof from thegroup consisting of halogen, cyano, C₁₋₆ alkyl, and C₁₋₆ alkoxy;

R⁵ is selected from the group consisting of C₁₋₆ alkyl, C₁₋₆ alkoxy,non-aromatic heterocycle, —NR⁶R⁷, and —CR⁸R⁹;

R⁶, R⁷, R⁸, and R⁹ are each independently selected from the groupconsisting of H, D, C₁₋₆ alkyl, and —(CH₂)_(k)OH;

or R⁶ and R⁷ are taken together with the nitrogen to which they areattached to form a morpholine ring;

or R⁸ and R⁹ are taken together with the carbon to which they areattached to form an oxetane ring;

R^(x) is independently selected at each occurrence thereof from thegroup consisting of H, D, —CH₂OC₁₋₆ alkyl, —(CH₂)_(m)C(O)NHR⁵,—(CH₂)_(m)C(O)NR⁶R⁷, and —CH₂C(O)R⁵ _(;)

R^(y) is independently selected at each occurrence thereof from thegroup consisting of H, D, C₁₋₆ alkyl, —CH₂OC₁₋₆ alkyl, —CH₂Ar, and—CH₂heteroaryl, wherein aryl (Ar) can be optionally substituted from 1to 3 times with a substituent selected independently at each occurrencethereof from the group consisting of halogen, cyano, C₁₋₆ alkyl, andC₁₋₆ alkoxy;

X is —(CH₂)_(q)—, —O—, or —(CD₂)_(q)—;

Y is O or S;

k is 1, 2, or 3;

m is 0, 1, 2, 3, 4, or 5;

n is 0, 1, 2, or 3;

p is 1 or 2;

q is 0, 1, or 2;

r is 1 or 2; and

s is 0 or 1;

with a proviso that when s is 0, then r is 2; and when s is 1, thenr+p≥3.

Another aspect of the present invention relates to a compound of Formula(Ia), Formula (Ib), or Formula (Ic):

wherein

R¹ is selected from the group consisting of monocyclic and bicyclicaryl, biphenyl, monocyclic and bicyclic heteroaryl, monocyclic andbicyclic heterocyclyl, and monocyclic and bicyclic non-aromaticheterocycle, wherein monocyclic and bicyclic aryl, biphenyl, monocyclicand bicyclic heteroaryl, monocyclic and bicyclic heterocyclyl, andmonocyclic and bicyclic non-aromatic heterocycle can be optionallysubstituted from 1 to 3 times with a substituent selected independentlyat each occurrence thereof from the group consisting of halogen, cyano,—CF₃, C₁₋₆ alkyl, and C₁₋₆ alkoxy;

R² is independently selected at each occurrence thereof from the groupconsisting of H, D, C₁₋₆ alkyl, —CH₂OC₁₋₆ alkyl, —CH₂Ar, and—CH₂heteroaryl, wherein aryl (Ar) can be optionally substituted from 1to 3 times with a substituent selected independently at each occurrencethereof from the group consisting of halogen, cyano, C₁₋₆ alkyl, andC₁₋₆ alkoxy;

R³ is independently selected at each occurrence thereof from the groupconsisting of H, D, —CH₂OC₁₋₆ alkyl, —(CH₂)_(m)C(O)NHR⁵, and—(CH₂)_(m)C(O)NR⁶R⁷ _(;)

R⁴ is selected from the group consisting of —C(O)(CH₂)_(n)Ph,—C(O)CH₂NR⁶R⁷, —SO₂Ar, —SO₂C₁₋₆ alkyl, —SO₂C₃₋₆ cycloalkyl,—C(O)(CH₂)_(n)Het, —C(O)C(O)Het, —C(O)C₁₋₆ alkyl, —C(O)OC₁₋₆ alkyl,—C(O)CF₃, heteroaryl, and —(CH₂)_(n)NR⁶R⁷, wherein aryl (Ar) andheteroaryl (Het) can be optionally substituted from 1 to 3 times with asubstituent selected independently at each occurrence thereof from thegroup consisting of halogen, cyano, C₁₋₆ alkyl, and C₁₋₆ alkoxy;

R⁵ is selected from the group consisting of C₁₋₆ alkyl, C₁₋₆ alkoxy,non-aromatic heterocycle, —NR⁶R⁷, and —CR⁸R⁹;

R⁶, R⁷, R⁸, and R⁹ are each independently selected from the groupconsisting of H, D, C₁₋₆ alkyl, and —(CH₂)_(k)OH;

or R⁶ and R⁷ are taken together with the nitrogen to which they areattached to form a morpholine ring;

or R⁸ and R⁹ are taken together with the carbon to which they areattached to form an oxetane ring;

R^(x) is independently selected at each occurrence thereof from thegroup consisting of H, D, —CH₂OC₁₋₆ alkyl, —(CH₂)_(m)C(O)NHR⁵,—(CH₂)_(m)C(O)NR⁶R⁷, and —CH₂C(O)R⁵ _(;)

R^(y) is independently selected at each occurrence thereof from thegroup consisting of H, D, C₁₋₆ alkyl, —CH₂OC₁₋₆ alkyl, —CH₂Ar, and—CH₂heteroaryl, wherein aryl (Ar) can be optionally substituted from 1to 3 times with a substituent selected independently at each occurrencethereof from the group consisting of halogen, cyano, C₁₋₆ alkyl, andC₁₋₆ alkoxy;

X is —(CH₂)_(q)—, —O—, or —(CD₂)_(q)—;

Y is O or S;

k is 1, 2, or 3;

m is 0, 1, 2, 3, 4, or 5;

n is 0, 1, 2, or 3;

q is 0, 1, or 2; and

s is 0 or 1;

or an oxide thereof, a pharmaceutically acceptable salt thereof, asolvate thereof, or a prodrug thereof.

Yet another aspect of the present invention relates to a method oftreating cancer, immunologic disorders, autoimmune disorders,neurodegenerative disorders, or inflammatory disorders in a subject orfor providing immunosuppression for transplanted organs or tissues in asubject. This method includes administering to the subject in needthereof a compound of the Formula (Ia), Formula (Ib), or Formula (Ic):

wherein

R¹ is selected from the group consisting of monocyclic and bicyclicaryl, biphenyl, monocyclic and bicyclic heteroaryl, monocyclic andbicyclic heterocyclyl, and monocyclic and bicyclic non-aromaticheterocycle, wherein monocyclic and bicyclic aryl, biphenyl, monocyclicand bicyclic heteroaryl, monocyclic and bicyclic heterocyclyl, andmonocyclic and bicyclic non-aromatic heterocycle can be optionallysubstituted from 1 to 3 times with a substituent selected independentlyat each occurrence thereof from the group consisting of halogen, cyano,—CF₃, C₁₋₆ alkyl, and C₁₋₆ alkoxy;

R² is independently selected at each occurrence thereof from the groupconsisting of H, D, C₁₋₆ alkyl, —CH₂OC₁₋₆ alkyl, —CH₂Ar, and—CH₂heteroaryl, wherein aryl (Ar) can be optionally substituted from 1to 3 times with a substituent selected independently at each occurrencethereof from the group consisting of halogen, cyano, C₁₋₆ alkyl, andC₁₋₆ alkoxy;

R³ is independently selected at each occurrence thereof from the groupconsisting of H, D, —CH₂OC₁₋₆ alkyl, —(CH₂)_(m)C(O)NHR⁵, and—(CH₂)_(m)C(O)NR⁶R⁷ _(;)

R⁴ is selected from the group consisting of —C(O)(CH₂)_(n)Ph,—C(O)CH₂NR⁶R⁷, —SO₂Ar, —SO₂C₁₋₆ alkyl, —SO₂C₃₋₆ cycloalkyl,—C(O)(CH₂)_(n)Het, —C(O)C(O)Het, —C(O)C₁₋₆ alkyl, —C(O)OC₁₋₆ alkyl,—C(O)CF₃, heteroaryl, and —(CH₂)_(n)NR⁶R⁷, wherein aryl (Ar) andheteroaryl (Het) can be optionally substituted from 1 to 3 times with asubstituent selected independently at each occurrence thereof from thegroup consisting of halogen, cyano, C₁₋₆ alkyl, and C₁₋₆ alkoxy;

R⁵ is selected from the group consisting of C₁₋₆ alkyl, C₁₋₆ alkoxy,non-aromatic heterocycle, —NR⁶R⁷, and —CR⁸R⁹;

R⁶, R⁷, R⁸, and R⁹ are each independently selected from the groupconsisting of H, D, C₁₋₆ alkyl, and —(CH₂)_(k)OH;

or R⁶ and R⁷ are taken together with the nitrogen to which they areattached to form a morpholine ring;

or R⁸ and R⁹ are taken together with the carbon to which they areattached to form an oxetane ring;

R^(x) is independently selected at each occurrence thereof from thegroup consisting of H, D, —CH₂OC₁₋₆ alkyl, —(CH₂)_(m)C(O)NHR⁵,—(CH₂)_(m)C(O)NR⁶R⁷, and —CH₂C(O)R⁵ _(;)

R^(y) is independently selected at each occurrence thereof from thegroup consisting of H, D, C₁₋₆ alkyl, —CH₂OC₁₋₆ alkyl, —CH₂Ar, and—CH₂heteroaryl, wherein aryl (Ar) can be optionally substituted from 1to 3 times with a substituent selected independently at each occurrencethereof from the group consisting of halogen, cyano, C₁₋₆ alkyl, andC₁₋₆ alkoxy;

X is —(CH₂)_(q)—, —O—, or —(CD₂)_(q)—;

Y is O or S;

k is 1, 2, or 3;

m is 0, 1, 2, 3, 4, or 5;

n is 0, 1, 2, or 3;

q is 0, 1, or 2; and

s is 0 or 1.

Selective inhibition of the i-20S is believed to impact the immunesystem but would otherwise be far less toxic than combined inhibition ofboth constitutive and immunoproteasomes. Here are presented inhibitorsthat act both with high selectivity and full reversibility on hu i-20Sβ5i over hu c-20S. Inhibitors that are selective for the i-20S β5i areexpected to be equally if not more efficacious in treating autoimmunedisease, with less toxicity. These inhibitors could open a new path tothe treatment of immunologic, autoimmune, inflammatory,neurodegenerative, and certain neoplastic disorders such as: systemiclupus erythematosus, chronic rheumatoid arthritis, inflammatory boweldisease, ulcerative colitis, Crohn's disease, multiple sclerosis,amyotrophic lateral sclerosis (ALS), atherosclerosis, scleroderma,systemic sclerosis, autoimmune hepatitis, Sjogren Syndrome, lupusnephritis, glomerulonephritis, rheumatoid arthritis, psoriasis,Myasthenia Gravis, Imunoglobuline A nephropathy, atherosclerosis,vasculitis, renal fibrosis, lung fibrosis, liver fibrosis, transplantrejection, idiopathic pulmonary fibrosis, asthma, and inflammationdriven cancers such as: triple negative breast cancers.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph showing inhibition of human i-20S β5i and c-20S β5c byselected dipeptidomimetics.

DETAILED DESCRIPTION OF THE INVENTION

One aspect of the present invention relates to a compound of Formula(I):

wherein

L is —(CR³R^(x))_(p)—;

M is —(CR²R^(y))_(r)—;

R¹ is selected from the group consisting of monocyclic and bicyclicaryl, biphenyl, monocyclic and bicyclic heteroaryl, monocyclic andbicyclic heterocyclyl, and monocyclic and bicyclic non-aromaticheterocycle, wherein monocyclic and bicyclic aryl, biphenyl, monocyclicand bicyclic heteroaryl, monocyclic and bicyclic heterocyclyl, andmonocyclic and bicyclic non-aromatic heterocycle can be optionallysubstituted from 1 to 3 times with a substituent selected independentlyat each occurrence thereof from the group consisting of halogen, cyano,—CF₃, C₁₋₆ alkyl, and C₁₋₆ alkoxy;

R² is independently selected at each occurrence thereof from the groupconsisting of H, D, C₁₋₆ alkyl, —CH₂OC₁₋₆ alkyl, —CH₂Ar, and—CH₂heteroaryl, wherein aryl (Ar) can be optionally substituted from 1to 3 times with a substituent selected independently at each occurrencethereof from the group consisting of halogen, cyano, C₁₋₆ alkyl, andC₁₋₆ alkoxy;

R³ is independently selected at each occurrence thereof from the groupconsisting of H, D, —CH₂OC₁₋₆ alkyl, —(CH₂)_(m)C(O)NHR⁵,—(CH₂)_(m)C(O)NR⁶R⁷, —(CH₂)_(m)C(O)OH, and —(CH₂)_(m)C(O)OBn;

R⁴ is selected from the group consisting of H, —C(O)(CH₂)Ph,—C(O)CH₂NR⁶R⁷, —SO₂Ar, —SO₂C₁₋₆ alkyl, —SO₂C₃₋₆ cycloalkyl,—C(O)(CH₂)_(n)Het, —C(O)C(O)Het, —C(O)C₁₋₆ alkyl, —C(O)OC₁₋₆ alkyl,—C(O)CF₃, heteroaryl, and —(CH₂)_(n)NR⁶R⁷, wherein aryl (Ar) andheteroaryl (Het) can be optionally substituted from 1 to 3 times with asubstituent selected independently at each occurrence thereof from thegroup consisting of halogen, cyano, C₁₋₆ alkyl, and C₁₋₆ alkoxy;

R⁵ is selected from the group consisting of C₁₋₆ alkyl, C₁₋₆ alkoxy,non-aromatic heterocycle, —NR⁶R⁷, and —CR⁸R⁹;

R⁶, R⁷, R⁸, and R⁹ are each independently selected from the groupconsisting of H, D, C₁₋₆ alkyl, and —(CH₂)_(k)OH;

or R⁶ and R⁷ are taken together with the nitrogen to which they areattached to form a morpholine ring;

or R⁸ and R⁹ are taken together with the carbon to which they areattached to form an oxetane ring;

R^(x) is independently selected at each occurrence thereof from thegroup consisting of H, D, —CH₂OC₁₋₆ alkyl, —(CH₂)_(m)C(O)NHR⁵,—(CH₂)_(m)C(O)NR⁶R⁷, and —CH₂C(O)R⁵ _(;)

R^(y) is independently selected at each occurrence thereof from thegroup consisting of H, D, C₁₋₆ alkyl, —CH₂OC₁₋₆ alkyl, —CH₂Ar, and—CH₂heteroaryl, wherein aryl (Ar) can be optionally substituted from 1to 3 times with a substituent selected independently at each occurrencethereof from the group consisting of halogen, cyano, C₁₋₆ alkyl, andC₁₋₆ alkoxy;

X is —(CH₂)_(q)—, —O—, or —(CD₂)_(q)—;

Y is O or S;

k is 1, 2, or 3;

m is 0, 1, 2, 3, 4, or 5;

n is 0, 1, 2, or 3;

p is 1 or 2;

q is 0, 1, or 2;

r is 1 or 2; and

s is 0 or 1;

with a proviso that when s is 0, then r is 2; and when s is 1, thenr+p≥3,

or an oxide thereof, a pharmaceutically acceptable salt thereof, asolvate thereof, or a prodrug thereof.

As used above, and throughout the description herein, the followingterms, unless otherwise indicated, shall be understood to have thefollowing meanings. If not defined otherwise herein, all technical andscientific terms used herein have the same meaning as is commonlyunderstood by one of ordinary skill in the art to which this technologybelongs. In the event that there is a plurality of definitions for aterm herein, those in this section prevail unless stated otherwise.

The term “alkyl” means an aliphatic hydrocarbon group which may bestraight or branched having about 1 to about 6 carbon atoms in thechain. Branched means that one or more lower alkyl groups such asmethyl, ethyl or propyl are attached to a linear alkyl chain. Exemplaryalkyl groups include methyl, ethyl, n-propyl, i-propyl, n-butyl,t-butyl, n-pentyl, and 3-pentyl.

The term “cycloalkyl” means a non-aromatic mono- or multicyclic ringsystem of about 3 to about 7 carbon atoms, preferably of about 5 toabout 7 carbon atoms. Exemplary monocyclic cycloalkyls includecyclopentyl, cyclohexyl, cycloheptyl, and the like.

The term “aryl” means an aromatic monocyclic or multicyclic ring systemof 6 to about 14 carbon atoms, preferably of 6 to about 10 carbon atoms.Representative aryl groups include phenyl and naphthyl.

The term “heteroaryl” means an aromatic monocyclic or multicyclic ringsystem of about 5 to about 14 ring atoms, preferably about 5 to about 10ring atoms, in which one or more of the atoms in the ring system is/areelement(s) other than carbon, for example, nitrogen, oxygen, or sulfur.In the case of multicyclic ring system, only one of the rings needs tobe aromatic for the ring system to be defined as “Heteroaryl”. Preferredheteroaryls contain about 5 to 6 ring atoms. The prefix aza, oxa, thia,or thio before heteroaryl means that at least a nitrogen, oxygen, orsulfur atom, respectively, is present as a ring atom. A nitrogen atom ofa heteroaryl is optionally oxidized to the corresponding N-oxide.Representative heteroaryls include pyridyl, 2-oxo-pyridinyl,pyrimidinyl, pyridazinyl, pyrazinyl, triazinyl, furanyl, pyrrolyl,thiophenyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl,isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, indolyl,isoindolyl, benzofuranyl, benzothiophenyl, indolinyl, 2-oxoindolinyl,dihydrobenzofuranyl, dihydrobenzothiophenyl, indazolyl, benzimidazolyl,benzooxazolyl, benzothiazolyl, benzoisoxazolyl, benzoisothiazolyl,benzotriazolyl, benzo[1,3]dioxolyl, quinolinyl, isoquinolinyl,quinazolinyl, cinnolinyl, pthalazinyl, quinoxalinyl,2,3-dihydro-benzo[1,4]dioxinyl, benzo[1,2,3]triazinyl,benzo[1,2,4]triazinyl, 4H-chromenyl, indolizinyl, quinolizinyl,6aH-thieno[2,3-d]imidazolyl, 1H-pyrrolo[2,3-b]pyridinyl,imidazo[1,2-a]pyridinyl, pyrazolo[1,5-a]pyridinyl,[1,2,4]triazolo[4,3-a]pyridinyl, [1,2,4]triazolo[1,5-a]pyridinyl,thieno[2,3-b]furanyl, thieno[2,3-b]pyridinyl, thieno[3,2-b]pyridinyl,furo[2,3-b]pyridinyl, furo[3,2-b]pyridinyl, thieno[3,2-d]pyrimidinyl,furo[3,2-d]pyrimidinyl, thieno[2,3-b]pyrazinyl, imidazo[1,2-a]pyrazinyl,5,6,7,8-tetrahydroimidazo[1,2-a]pyrazinyl,6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazinyl,2-oxo-2,3-dihydrobenzo[d]oxazolyl, 3,3-dimethyl-2-oxoindolinyl,2-oxo-2,3-dihydro-1H-pyrrolo[2,3-b]pyridinyl,benzo[c][1,2,5]oxadiazolyl, benzo[c][1,2,5]thiadiazolyl,3,4-dihydro-2H-benzo[b][1,4]oxazinyl,5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazinyl,[1,2,4]triazolo[4,3-a]pyrazinyl,3-oxo-[1,2,4]triazolo[4,3-a]pyridin-2(3H)-yl, and the like.

As used herein, “heterocyclyl” or “heterocycle” refers to a stable 3- to18-membered ring (radical) which consists of carbon atoms and from oneto five heteroatoms selected from the group consisting of nitrogen,oxygen and sulfur. For purposes of this application, the heterocycle maybe a monocyclic, or a polycyclic ring system, which may include fused,bridged, or spiro ring systems; and the nitrogen, carbon, or sulfuratoms in the heterocycle may be optionally oxidized; the nitrogen atommay be optionally quaternized; and the ring may be partially or fullysaturated. Examples of such heterocycles include, without limitation,azepinyl, azocanyl, pyranyl dioxanyl, dithianyl, 1,3-dioxolanyl,tetrahydrofuryl, dihydropyrrolidinyl, decahydroisoquinolyl,imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl,octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl,2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazepinyl, oxazolidinyl,oxiranyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl,pyrazolidinyl, thiazolidinyl, tetrahydropyranyl, thiamorpholinyl,thiamorpholinyl sulfoxide, and thiamorpholinyl sulfone. Furtherheterocycles and heteroaryls are described in Katritzky et al., eds.,Comprehensive Heterocyclic Chemistry: The Structure, Reactions,Synthesis and Use of Heterocyclic Compounds, Vol. 1-8, Pergamon Press,N.Y. (1984), which is hereby incorporated by reference in its entirety.

The term “non-aromatic heterocycle” means a non-aromatic monocyclicsystem containing 3 to 10 atoms, preferably 4 to about 7 carbon atoms,in which one or more of the atoms in the ring system is/are element(s)other than carbon, for example, nitrogen, oxygen, or sulfur.Representative non-aromatic heterocycle groups include pyrrolidinyl,2-oxopyrrolidinyl, piperidinyl, 2-oxopiperidinyl, azepanyl,2-oxoazepanyl, 2-oxooxazolidinyl, morpholino, 3-oxomorpholino,thiomorpholino, 1,1-dioxothiomorpholino, piperazinyl,tetrahydro-2H-oxazinyl, and the like.

The term “monocyclic” used herein indicates a molecular structure havingone ring.

The term “polycyclic” or “multi-cyclic” used herein indicates amolecular structure having two or more rings, including, but not limitedto, fused, bridged, or spiro rings.

Terminology related to “protecting”, “deprotecting,” and “protected”functionalities occurs throughout this application. Such terminology iswell understood by persons of skill in the art and is used in thecontext of processes which involve sequential treatment with a series ofreagents. In that context, a protecting group refers to a group which isused to mask a functionality during a process step in which it wouldotherwise react, but in which reaction is undesirable. The protectinggroup prevents reaction at that step, but may be subsequently removed toexpose the original functionality. The removal or “deprotection” occursafter the completion of the reaction or reactions in which thefunctionality would interfere. Thus, when a sequence of reagents isspecified, as it is in the processes described herein, the person ofordinary skill can readily envision those groups that would be suitableas “protecting groups.” Suitable groups for that purpose are discussedin standard textbooks in the field of chemistry, such as Greene,Protective Groups in Organic Synthesis, John Wiley & Sons, New York(1991), which is hereby incorporated by reference in its entirety.

The term “alkoxy” means groups of from 1 to 8 carbon atoms of astraight, branched, or cyclic configuration and combinations thereofattached to the parent structure through an oxygen. Examples includemethoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy, cyclohexyloxy, andthe like. Lower-alkoxy refers to groups containing one to four carbons.For the purposes of the present patent application, alkoxy also includesmethylenedioxy and ethylenedioxy in which each oxygen atom is bonded tothe atom, chain, or ring from which the methylenedioxy or ethylenedioxygroup is pendant so as to form a ring. Thus, for example, phenylsubstituted by alkoxy may be, for example,

A compound with a hydroxy group drawn next to a nitrogen on aheterocycle can exist as the “keto” form. For example,3-(2-hydroxy-[1,2,4]triazolo[1,5-a]pyridin-6-yl)propanoic acid can existas 3-(2-oxo-2,3-dihydro-[1,2,4]triazolo[1,5-a]pyridin-6-yl)propanoicacid.

The term “halo” or “halogen” means fluoro, chloro, bromo, or iodo.

The term “substituted” or “substitution” of an atom means that one ormore hydrogen on the designated atom is replaced with a selection fromthe indicated group, provided that the designated atom's normal valencyis not exceeded.

“Unsubstituted” atoms bear all of the hydrogen atoms dictated by theirvalency. When a substituent is keto (i.e., =0), then two hydrogens onthe atom are replaced. Combinations of substituents and/or variables arepermissible only if such combinations result in stable compounds; by“stable compound” or “stable structure” is meant a compound that issufficiently robust to survive isolation to a useful degree of purityfrom a reaction mixture, and formulation into an efficacious therapeuticagent.

The term “optionally substituted” is used to indicate that a group mayhave a substituent at each substitutable atom of the group (includingmore than one substituent on a single atom), provided that thedesignated atom's normal valency is not exceeded and the identity ofeach substituent is independent of the others. Up to three H atoms ineach residue are replaced with alkyl, halogen, haloalkyl, hydroxy, loweralkoxy, carboxy, carboalkoxy (also referred to as alkoxycarbonyl),carboxamido (also referred to as alkylaminocarbonyl), cyano, carbonyl,nitro, amino, alkylamino, dialkylamino, mercapto, alkylthio, sulfoxide,sulfone, acylamino, amidino, phenyl, benzyl, heteroaryl, phenoxy,benzyloxy, or heteroaryloxy. “Unsubstituted” atoms bear all of thehydrogen atoms dictated by their valency. When a substituent is keto(i.e., =0), then two hydrogens on the atom are replaced. Combinations ofsubstituents and/or variables are permissible only if such combinationsresult in stable compounds; by “stable compound” or “stable structure”is meant a compound that is sufficiently robust to survive isolation toa useful degree of purity from a reaction mixture, and formulation intoan efficacious therapeutic agent.

The term “method of treating” means amelioration or relief from thesymptoms and/or effects associated with the disorders described herein.As used herein, reference to “treatment” of a patient is intended toinclude prophylaxis.

The term “compounds of the invention”, and equivalent expressions, aremeant to embrace compounds of general formula (I), formula (Ia), formula(Ib), and formula (Ic) as hereinbefore described, which expressionincludes the prodrugs, the pharmaceutically acceptable salts, and thesolvates, e.g. hydrates, where the context so permits. Similarly,reference to intermediates, whether or not they themselves are claimed,is meant to embrace their salts, and solvates, where the context sopermits. For the sake of clarity, particular instances when the contextso permits are sometimes indicated in the text, but these instances arepurely illustrative and it is not intended to exclude other instanceswhen the context so permits.

The term “pharmaceutically acceptable salts” means the relativelynon-toxic, inorganic, and organic acid addition salts, and base additionsalts, of compounds of the present invention. These salts can beprepared in situ during the final isolation and purification of thecompounds. In particular, acid addition salts can be prepared byseparately reacting the purified compound in its free base form with asuitable organic or inorganic acid and isolating the salt thus formed.Exemplary acid addition salts include the hydrobromide, hydrochloride,sulfate, bisulfate, phosphate, nitrate, acetate, oxalate, valerate,oleate, palmitate, stearate, laurate, borate, benzoate, lactate,phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate,naphthylate, mesylate, glucoheptonate, lactiobionate, sulphamates,malonates, salicylates, propionates, methylene-bis-b-hydroxynaphthoates,gentisates, isethionates, di-p-toluoyltartrates, methanesulphonates,ethanesulphonates, benzenesulphonates, p-toluenesulphonates,cyclohexylsulphamates and quinateslaurylsulphonate salts, and the like(see, for example, Berge et al., “Pharmaceutical Salts,” J. Pharm. Sci.,66:1-9 (1977) and Remington's Pharmaceutical Sciences, 17th ed., MackPublishing Company, Easton, Pa., 1985, p. 1418, which are herebyincorporated by reference in their entirety). Base addition salts canalso be prepared by separately reacting the purified compound in itsacid form with a suitable organic or inorganic base and isolating thesalt thus formed. Base addition salts include pharmaceuticallyacceptable metal and amine salts. Suitable metal salts include thesodium, potassium, calcium, barium, zinc, magnesium, and aluminum salts.The sodium and potassium salts are preferred. Suitable inorganic baseaddition salts are prepared from metal bases which include, for example,sodium hydride, sodium hydroxide, potassium hydroxide, calciumhydroxide, aluminium hydroxide, lithium hydroxide, magnesium hydroxide,and zinc hydroxide. Suitable amine base addition salts are prepared fromamines which have sufficient basicity to form a stable salt, andpreferably include those amines which are frequently used in medicinalchemistry because of their low toxicity and acceptability for medicaluse, such as ammonia, ethylenediamine, N-methyl-glutamine, lysine,arginine, ornithine, choline, N,N′-dibenzylethylenediamine,chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine,diethylamine, piperazine, tris(hydroxymethyl)-aminomethane,tetramethylammonium hydroxide, triethylamine, dibenzylamine, ephenamine,dehydroabietylamine, N-ethylpiperidine, benzylamine,tetramethylammonium, tetraethylammonium, methylamine, dimethylamine,trimethylamine, ethylamine, basic amino acids, e.g., lysine andarginine, dicyclohexylamine, and the like.

The term “pharmaceutically acceptable prodrugs” as used herein meansthose prodrugs of the compounds useful according to the presentinvention which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswith undue toxicity, irritation, allergic response, and the like,commensurate with a reasonable benefit/risk ratio, and effective fortheir intended use, as well as the zwitterionic forms, where possible,of the compounds of the invention. The term “prodrug” means compoundsthat are rapidly transformed in vivo to yield the parent compound of theabove formula, for example by hydrolysis in blood. Functional groupswhich may be rapidly transformed, by metabolic cleavage, in vivo form aclass of groups reactive with the carboxyl group of the compounds ofthis invention. They include, but are not limited to, such groups asalkanoyl (such as acetyl, propionyl, butyryl, and the like),unsubstituted and substituted aroyl (such as benzoyl and substitutedbenzoyl), alkoxycarbonyl (such as ethoxycarbonyl), trialkylsilyl (suchas trimethyl- and triethysilyl), monoesters formed with dicarboxylicacids (such as succinyl), and the like. Because of the ease with whichthe metabolically cleavable groups of the compounds useful according tothis invention are cleaved in vivo, the compounds bearing such groupsact as pro-drugs. The compounds bearing the metabolically cleavablegroups have the advantage that they may exhibit improved bioavailabilityas a result of enhanced solubility and/or rate of absorption conferredupon the parent compound by virtue of the presence of the metabolicallycleavable group. A thorough discussion of prodrugs is provided in thefollowing: Design of Prodrugs, H. Bundgaard, ed., Elsevier (1985);Methods in Enzymology, K. Widder et al, Ed., Academic Press, 42, p.309-396 (1985); A Textbook of Drug Design and Development,Krogsgaard-Larsen and H. Bundgaard, ed., Chapter 5; “Design andApplications of Prodrugs” p. 113-191 (1991); Advanced Drug DeliveryReviews, H. Bundgard, 8, p. 1-38 (1992); J. Pharm. Sci., 77:285 (1988);Nakeya et al, Chem. Pharm. Bull., 32:692 (1984); Higuchi et al.,“Pro-drugs as Novel Delivery Systems,” Vol. 14 of the A.C.S. SymposiumSeries, and Bioreversible Carriers in Drug Design, Edward B. Roche, ed.,American Pharmaceutical Association and Pergamon Press (1987), which areincorporated herein by reference in their entirety. Examples of prodrugsinclude, but are not limited to, acetate, formate, and benzoatederivatives of alcohol and amine functional groups in the compounds ofthe invention.

The term “solvate” refers to a compound of Formula (I), Formula (Ia),Formula (Ib), and Formula (Ic) in the solid state, wherein molecules ofa suitable solvent are incorporated in the crystal lattice. A suitablesolvent for therapeutic administration is physiologically tolerable atthe dosage administered. Examples of suitable solvents for therapeuticadministration are ethanol and water. When water is the solvent, thesolvate is referred to as a hydrate. In general, solvates are formed bydissolving the compound in the appropriate solvent and isolating thesolvate by cooling or using an antisolvent. The solvate is typicallydried or azeotroped under ambient conditions.

The term “therapeutically effective amounts” is meant to describe anamount of compound of the present invention effective to produce thedesired therapeutic effect. Such amounts generally vary according to anumber of factors well within the purview of ordinarily skilled artisansgiven the description provided herein to determine and account for.These include, without limitation: the particular subject, as well asits age, weight, height, general physical condition, and medicalhistory; the particular compound used, as well as the carrier in whichit is formulated and the route of administration selected for it; and,the nature and severity of the condition being treated.

The term “pharmaceutical composition” means a composition comprising acompound of Formula (I) Formula (Ia), Formula (Ib), and Formula (Ic) andat least one component comprising pharmaceutically acceptable carriers,diluents, adjuvants, excipients, or vehicles, such as preserving agents,fillers, disintegrating agents, wetting agents, emulsifying agents,suspending agents, sweetening agents, flavoring agents, perfumingagents, antibacterial agents, antifungal agents, lubricating agents anddispensing agents, depending on the nature of the mode of administrationand dosage forms. Examples of suspending agents include ethoxylatedisostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters,microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agarand tragacanth, or mixtures of these substances. Prevention of theaction of microorganisms can be ensured by various antibacterial andantifungal agents, for example, parabens, chlorobutanol, phenol, sorbicacid, and the like. It may also be desirable to include isotonic agents,for example sugars, sodium chloride, and the like. Prolonged absorptionof the injectable pharmaceutical form can be brought about by the use ofagents delaying absorption, for example, aluminum monostearate andgelatin. Examples of suitable carriers, diluents, solvents, or vehiclesinclude water, ethanol, polyols, suitable mixtures thereof, vegetableoils (such as olive oil), and injectable organic esters such as ethyloleate. Examples of excipients include lactose, milk sugar, sodiumcitrate, calcium carbonate, and dicalcium phosphate. Examples ofdisintegrating agents include starch, alginic acids, and certain complexsilicates. Examples of lubricants include magnesium stearate, sodiumlauryl sulphate, talc, as well as high molecular weight polyethyleneglycols.

The term “pharmaceutically acceptable” means it is, within the scope ofsound medical judgement, suitable for use in contact with the cells ofhumans and lower animals without undue toxicity, irritation, allergicresponse and the like, and are commensurate with a reasonablebenefit/risk ratio.

The term “pharmaceutically acceptable dosage forms” means dosage formsof the compound of the invention, and includes, for example, tablets,dragees, powders, elixirs, syrups, liquid preparations, includingsuspensions, sprays, inhalants tablets, lozenges, emulsions, solutions,granules, capsules, and suppositories, as well as liquid preparationsfor injections, including liposome preparations. Techniques andformulations generally may be found in Remington's PharmaceuticalSciences, Mack Publishing Co., Easton, Pa., latest edition.

Compounds described herein may contain one or more asymmetric centersand may thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms. Each chiral center may be defined, in terms ofabsolute stereochemistry, as (R)- or (S)-. This technology is meant toinclude all such possible isomers, as well as mixtures thereof,including racemic and optically pure forms. Optically active (R)- and(S)-, (−)- and (+)-, or (D)- and (L)-isomers may be prepared usingchiral synthons or chiral reagents, or resolved using conventionaltechniques. When the compounds described herein contain olefinic doublebonds or other centers of geometric asymmetry, and unless specifiedotherwise, it is intended that the compounds include both E and Zgeometric isomers. Likewise, all tautomeric forms are also intended tobe included.

This technology also envisions the “quaternization” of any basicnitrogen-containing groups of the compounds disclosed herein. The basicnitrogen can be quaternized with any agents known to those of ordinaryskill in the art including, for example, lower alkyl halides, such asmethyl, ethyl, propyl and butyl chloride, bromides and iodides; dialkylsulfates including dimethyl, diethyl, dibutyl and diamyl sulfates; longchain halides such as decyl, lauryl, myristyl and stearyl chlorides,bromides and iodides; and aralkyl halides including benzyl and phenethylbromides. Water or oil-soluble or dispersible products may be obtainedby such quaternization.

In the characterization of some of the substituents, it is recited thatcertain substituents may combine to form rings. Unless stated otherwise,it is intended that such rings may exhibit various degrees ofunsaturation (from fully saturated to fully unsaturated), may includeheteroatoms and may be substituted with lower alkyl or alkoxy.

Compounds of the present invention can be produced according to knownmethods. For example, compounds of the present invention wherein s is 0can be prepared according to Scheme 1 and Scheme 2 outlined below.

Reaction of the carboxylic acid derivative (1) with amine R¹X—NH₂ (2)leads to formation of the compound (3). The reaction can be carried outin a variety of solvents, for example in methylene chloride (CH₂Cl₂),tetrahydrofuran (THF), dimethylformamide (DMF), or other such solventsor in the mixture of such solvents. During the reaction process, thenon-participating carboxylic acids or amines on the reacting set ofamino acids or peptide fragments can be protected by a suitableprotecting group which can be selectively removed at a later time ifdesired. A detailed description of these groups and their selection andchemistry is contained in “The Peptides, Vol, 3”, Gross and Meinenhofer,Eds., Academic Press, New York, 1981, which is hereby incorporated byreference in its entirety. Thus, useful protective groups for the aminogroup are benzyloxycarbonyl (Cbz), t-butyloxycarbonyl (t-BOC),2,2,2-trichloroethoxycarbonyl (Troc), t-amyloxycarbonyl,4-methoxybenzyloxycarbonyl, 2-(trichlorosilyl)ethoxycarbonyl,9-fluorenylmethoxycarbonyl (Fmoc), phthaloyl, acetyl (Ac), formyl,trifluoroacetyl, and the like. Following the deprotection, amine (4) isreacted with R⁴-LG (5) (wherein LG is a suitable leaving group) to formfinal product (6).

Alternatively, carboxylic acid derivative bearing protecting group (PG)(1) can be first deprotected and then reacted with the R⁴-LG (5)(wherein LG is a suitable leaving group) to form compound (8). Compound(8) can be then reacted with amine (2), R¹X—NH₂, to form final product(6).

Compounds of the present invention wherein s is 1 can be preparedaccording to the general schemes outlined below (Schemes 3-7).

The compounds of the present invention may be prepared by stepwisecoupling of the amino acids. The coupling reactions are conducted insolvents such as methylene chloride (CH₂Cl₂), tetrahydrofuran (THF),dimethylformamide (DMF), or other such solvents. During the couplingprocess, the non-participating carboxylic acids or amines on thereacting set of amino acids or peptide fragments can be protected by asuitable protecting group which can be selectively removed at a latertime if desired. A detailed description of these groups and theirselection and chemistry is contained in “The Peptides, Vol. 3”, Grossand Meinenhofer, Eds., Academic Press, New York, 1981, which is herebyincorporated by reference in its entirety. Thus, useful protectivegroups for the amino group are benzyloxycarbonyl (Cbz),t-butyloxycarbonyl (t-BOC), 2,2,2-trichloroethoxycarbonyl (Troc),t-amyloxycarbonyl, 4-methoxybenzyloxycarbonyl,2-(trichlorosilyl)ethoxycarbonyl, 9-fluorenylmethoxycarbonyl (Fmoc),phthaloyl, acetyl (Ac), formyl, trifluoroacetyl, and the like.Carboxylic acid bearing protecting group (PG) (9) is coupled with theamine (4) to form compound (10). Following the deprotection reaction,compound (11) is reacted with R⁴-LG (5) to form final product (12).

Alternatively, carboxylic acid bearing protecting group (PG) (13) can befirst reacted with R⁴-LG (wherein LG is a suitable leaving group) (5) toform compound (14). Then, following the deprotection reaction, acid (15)can be coupled with amine (4) to form final product (12).

Alternatively, acid (15) can be first coupled with amino acid (16) toform compound (17). Following the deprotection reaction, acid (18) canbe reacted with amine (2), R¹X—NH₂, to form final product (12).

Alternatively, amino acid (9a) can be coupled with amino acid (16a) toform compound (19). Following the deprotection reaction, acid (20) canbe reacted with R⁴-LG (wherein LG is a suitable leaving group) (5) toform compound (21). Following the deprotection reaction, acid (22) canbe coupled with amine (2), R¹X—NH₂, to form final product (12).

Alternatively, following the deprotection reaction, acid (23) can becoupled with amine (2), R¹X—NH₂, to form compound (10a). Following thedeprotection reaction, amine (11) can then be reacted with R⁴-LG(wherein LG is a suitable leaving group) (5) to form final product (12).

Compounds of the present invention wherein Y is O can be easilyconverted to their analogs wherein Y is S, according to known methods.For example by using Lawesson's reagent.

Another aspect of the present invention relates to a compound of Formula(Ia), Formula (Ib), or Formula (Ic):

wherein

R¹ is selected from the group consisting of monocyclic and bicyclicaryl, biphenyl, monocyclic and bicyclic heteroaryl, monocyclic andbicyclic heterocyclyl, and monocyclic and bicyclic non-aromaticheterocycle, wherein monocyclic and bicyclic aryl, biphenyl, monocyclicand bicyclic heteroaryl, monocyclic and bicyclic heterocyclyl, andmonocyclic and bicyclic non-aromatic heterocycle can be optionallysubstituted from 1 to 3 times with a substituent selected independentlyat each occurrence thereof from the group consisting of halogen, cyano,—CF₃, C₁₋₆ alkyl, and C₁₋₆ alkoxy;

R² is independently selected at each occurrence thereof from the groupconsisting of H, D, C₁₋₆ alkyl, —CH₂OC₁₋₆ alkyl, —CH₂Ar, and—CH₂heteroaryl, wherein aryl (Ar) can be optionally substituted from 1to 3 times with a substituent selected independently at each occurrencethereof from the group consisting of halogen, cyano, C₁₋₆ alkyl, andC₁₋₆ alkoxy;

R³ is independently selected at each occurrence thereof from the groupconsisting of H, D, —CH₂OC₁₋₆ alkyl, —(CH₂)_(m)C(O)NHR⁵, and—(CH₂)_(m)C(O)NR⁶R⁷ _(;)

R⁴ is selected from the group consisting of —C(O)(CH₂)_(n)Ph,—C(O)CH₂NR⁶R⁷, —SO₂Ar, —SO₂C₁₋₆ alkyl, —SO₂C₃₋₆ cycloalkyl,—C(O)(CH₂)_(n)Het, —C(O)C(O)Het, —C(O)C₁₋₆ alkyl, —C(O)OC₁₋₆ alkyl,—C(O)CF₃, heteroaryl, and —(CH₂)_(n)NR⁶R⁷, wherein aryl (Ar) andheteroaryl (Het) can be optionally substituted from 1 to 3 times with asubstituent selected independently at each occurrence thereof from thegroup consisting of halogen, cyano, C₁₋₆ alkyl, and C₁₋₆ alkoxy;

R⁵ is selected from the group consisting of C₁₋₆ alkyl, C₁₋₆ alkoxy,non-aromatic heterocycle, —NR⁶R⁷, and —CR⁸R⁹;

R⁶, R⁷, R⁸, and R⁹ are each independently selected from the groupconsisting of H, D, C₁₋₆ alkyl, and —(CH₂)_(k)OH;

or R⁶ and R⁷ are taken together with the nitrogen to which they areattached to form a morpholine ring;

or R⁸ and R⁹ are taken together with the carbon to which they areattached to form an oxetane ring;

R^(x) is independently selected at each occurrence thereof from thegroup consisting of H, D, —CH₂OC₁₋₆ alkyl, —(CH₂)_(m)C(O)NHR⁵,—(CH₂)_(m)C(O)NR⁶R⁷, and —CH₂C(O)R⁵ _(;)

R^(y) is independently selected at each occurrence thereof from thegroup consisting of H, D, C₁₋₆ alkyl, —CH₂OC₁₋₆ alkyl, —CH₂Ar, and—CH₂heteroaryl, wherein aryl (Ar) can be optionally substituted from 1to 3 times with a substituent selected independently at each occurrencethereof from the group consisting of halogen, cyano, C₁₋₆ alkyl, andC₁₋₆ alkoxy;

X is —(CH₂)_(q)—, —O—, or —(CD₂)_(q)—;

Y is O or S;

k is 1, 2, or 3;

m is 0, 1, 2, 3, 4, or 5;

n is 0, 1, 2, or 3;

q is 0, 1, or 2; and

s is 0 or 1;

or an oxide thereof, a pharmaceutically acceptable salt thereof, asolvate thereof, or a prodrug thereof.

Compounds according to the present invention include compounds ofFormula (I), Formula (Ia), Formula (Ib), and Formula (Ic). In oneembodiment compound according to the present invention is a compound ofFormula (I). In another embodiment the compound according to the presentinvention is a compound of Formula (Ia), Formula (Ib), or Formula (Ic).

In one embodiment, compound has the formula:

In another embodiment, compound has the formula

One embodiment relates to the compounds of the present invention whereR¹ is selected from the group consisting of

and

R¹¹ is selected from the group consisting of halogen, cyano, —CF₃, C₁₋₆alkyl, and C₁₋₆ alkoxy.

Another embodiment relates to the compounds of the present inventionwhere R² is selected from the group consisting of H, Me, —CH₂(Me)₂,—CH₂OMe,

Another embodiment relates to the compounds of the present inventionwhere R² is selected from the group consisting of Me, —CH₂(Me)₂,—CH₂OMe,

One embodiment relates to the compounds of the present invention whereR³ is selected from the group consisting of —CH₂OMe, —CH₂C(O)OH,—CH₂C(O)OBn, —(CH₂)₂C(O)OBn, —(CH₂)₂C(O)OH,

Another embodiments relates to the compounds of the present inventionwhere R³ is selected from the group consisting of —CH₂OMe,

Another embodiment relates to the compounds of the present inventionwhere R⁴ is selected from the group consisting of H, trifluoroacetyl,

l is 0, 1, 2, 3, or 4;

m is 0, 1, 2, 3, 4, or 5;

n is 0, 1, 2, or 3; and

R is selected from the group consisting of H, halogen, cyano, C₁₋₆alkyl, and C₁₋₆ alkoxy.

Yet another embodiment relates to the compounds of the present inventionwhere R⁴ is selected from the group consisting of trifluoroacetyl,

l is 0, 1, 2, 3, or 4;

m is 0, 1, 2, 3, 4, or 5;

n is 0, 1, 2, or 3; and

R is selected from the group consisting of H, halogen, cyano, C₁₋₆alkyl, and C₁₋₆ alkoxy.

One embodiment relates to the compounds of the present invention wherethe compound has a structure selected from the group consisting of:

Another embodiments relates to the compound of the present inventionwhere the compound has a structure selected from the group consistingof:

While it may be possible for compounds of the present invention to beadministered as raw chemicals, it will often be preferable to presentthem as a part of a pharmaceutical composition. Accordingly, anotheraspect of the present invention is a pharmaceutical compositioncontaining a therapeutically effective amount of the compound of thepresent invention, or a pharmaceutically acceptable salt or solvatethereof, and a pharmaceutically acceptable carrier. The carrier must be“acceptable” in the sense of being compatible with the other ingredientsof the formulation and not deleterious to the recipient thereof.

Compounds according to the present invention include compounds ofFormula (I), Formula (Ia), Formula (Ib), and Formula (Ic). In oneembodiment compound according to the present invention is a compound ofFormula (I). In another embodiment the compound according to the presentinvention is a compound of Formula (Ia), Formula (Ib), or Formula (Ic).

In practicing the method of the present invention, agents suitable fortreating a subject can be administered using any method standard in theart. The agents, in their appropriate delivery form, can be administeredorally, intradermally, intramuscularly, intraperitoneally,intravenously, subcutaneously, or intranasally. The compositions of thepresent invention may be administered alone or with suitablepharmaceutical carriers, and can be in solid or liquid form, such astablets, capsules, powders, solutions, suspensions, or emulsions.

The agents of the present invention may be orally administered, forexample, with an inert diluent, or with an assimilable edible carrier,or it may be enclosed in hard or soft shell capsules, or it may becompressed into tablets, or they may be incorporated directly with thefood of the diet. Agents of the present invention may also beadministered in a time release manner incorporated within such devicesas time-release capsules or nanotubes. Such devices afford flexibilityrelative to time and dosage. For oral therapeutic administration, theagents of the present invention may be incorporated with excipients andused in the form of tablets, capsules, elixirs, suspensions, syrups, andthe like. Such compositions and preparations should contain at least0.1% of the agent, although lower concentrations may be effective andindeed optimal. The percentage of the agent in these compositions may,of course, be varied and may conveniently be between about 2% to about60% of the weight of the unit. The amount of an agent of the presentinvention in such therapeutically useful compositions is such that asuitable dosage will be obtained.

Also specifically contemplated are oral dosage forms of the agents ofthe present invention. The agents may be chemically modified so thatoral delivery of the derivative is efficacious. Generally, the chemicalmodification contemplated is the attachment of at least one moiety tothe component molecule itself, where said moiety permits (a) inhibitionof proteolysis; and (b) uptake into the blood stream from the stomach orintestine. Also desired is the increase in overall stability of thecomponent or components and increase in circulation time in the body.Examples of such moieties include: polyethylene glycol, copolymers ofethylene glycol and propylene glycol, carboxymethyl cellulose, dextran,polyvinyl alcohol, polyvinyl pyrrolidone and polyproline. (Abuchowskiand Davis, “Soluble Polymer-Enzyme Adducts,” In: Enzymes as Drugs,Hocenberg and Roberts, eds., Wiley-Interscience, New York, N.Y., pp.367-383 (1981), which are hereby incorporated by reference in theirentirety). Other polymers that could be used are poly-1,3-dioxolane andpoly-1,3,6-tioxocane. Preferred for pharmaceutical usage, as indicatedabove, are polyethylene glycol moieties.

The tablets, capsules, and the like may also contain a binder such asgum tragacanth, acacia, corn starch, or gelatin; excipients such asdicalcium phosphate; a disintegrating agent such as corn starch, potatostarch, alginic acid; a lubricant such as magnesium stearate; and asweetening agent such as sucrose, lactose, sucralose, or saccharin. Whenthe dosage unit form is a capsule, it may contain, in addition tomaterials of the above type, a liquid carrier such as a fatty oil.

Various other materials may be present as coatings or to modify thephysical form of the dosage unit. For instance, tablets may be coatedwith shellac, sugar, or both. A syrup may contain, in addition to activeingredient, sucrose as a sweetening agent, methyl and propylparabens aspreservatives, a dye, and flavoring such as cherry or orange flavor.

The agents of the present invention may also be administeredparenterally. Solutions or suspensions of the agent can be prepared inwater suitably mixed with a surfactant such as hydroxypropylcellulose.Dispersions can also be prepared in glycerol, liquid polyethyleneglycols, and mixtures thereof in oils. Illustrative oils are those ofpetroleum, animal, vegetable, or synthetic origin, for example, peanutoil, soybean oil, or mineral oil. In general, water, saline, aqueousdextrose and related sugar solution, and glycols, such as propyleneglycol or polyethylene glycol, are preferred liquid carriers,particularly for injectable solutions. Under ordinary conditions ofstorage and use, these preparations contain a preservative to preventthe growth of microorganisms.

The pharmaceutical forms suitable for injectable use include sterileaqueous solutions or dispersions and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions. In all cases, the form must be sterile and must be fluid tothe extent that easy syringability exists. It must be stable under theconditions of manufacture and storage and must be preserved against thecontaminating action of microorganisms, such as bacteria and fungi. Thecarrier can be a solvent or dispersion medium containing, for example,water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquidpolyethylene glycol), suitable mixtures thereof, and vegetable oils.

When it is desirable to deliver the agents of the present inventionsystemically, they may be formulated for parenteral administration byinjection, e.g., by bolus injection or continuous infusion. Formulationsfor injection may be presented in unit dosage form, e.g., in ampoules orin multi-dose containers, with an added preservative. The compositionsmay take such forms as suspensions, solutions or emulsions in oily oraqueous vehicles, and may contain formulatory agents such as suspending,stabilizing and/or dispersing agents.

Intraperitoneal or intrathecal administration of the agents of thepresent invention can also be achieved using infusion pump devices suchas those described by Medtronic, Northridge, Calif. Such devices allowcontinuous infusion of desired compounds avoiding multiple injectionsand multiple manipulations.

In addition to the formulations described previously, the agents mayalso be formulated as a depot preparation. Such long acting formulationsmay be formulated with suitable polymeric or hydrophobic materials (forexample as an emulsion in an acceptable oil) or ion exchange resins, oras sparingly soluble derivatives, for example, as a sparingly solublesalt.

The agents of the present invention may also be administered directly tothe airways in the form of an aerosol. For use as aerosols, the agent ofthe present invention in solution or suspension may be packaged in apressurized aerosol container together with suitable propellants, forexample, hydrocarbon propellants like propane, butane, or isobutane withconventional adjuvants. The agent of the present invention also may beadministered in a non-pressurized form such as in a nebulizer oratomizer.

Effective doses of the compositions of the present invention, for thetreatment of cancer or pathogen infection vary depending upon manydifferent factors, including type and stage of cancer or the type ofpathogen infection, means of administration, target site, physiologicalstate of the patient, other medications or therapies administered, andphysical state of the patient relative to other medical complications.Treatment dosages need to be titrated to optimize safety and efficacy.

The percentage of active ingredient in the compositions of the presentinvention may be varied, it being necessary that it should constitute aproportion such that a suitable dosage shall be obtained. Obviously,several unit dosage forms may be administered at about the same time.The dose employed will be determined by the physician, and depends uponthe desired therapeutic effect, the route of administration and theduration of the treatment, and the condition of the patient. In theadult, the doses are generally from about 0.01 to about 100 mg/kg bodyweight, preferably about 0.01 to about 10 mg/kg body weight per day byinhalation, from about 0.01 to about 100 mg/kg body weight, preferably0.1 to 70 mg/kg body weight, more especially 0.1 to 10 mg/kg body weightper day by oral administration, and from about 0.01 to about 50 mg/kgbody weight, preferably 0.01 to 10 mg/kg body weight per day byintravenous administration. In each particular case, the doses will bedetermined in accordance with the factors distinctive to the subject tobe treated, such as age, weight, general state of health, and othercharacteristics which can influence the efficacy of the medicinalproduct.

The products according to the present invention may be administered asfrequently as necessary in order to obtain the desired therapeuticeffect. Some patients may respond rapidly to a higher or lower dose andmay find much weaker maintenance doses adequate. For other patients, itmay be necessary to have long-term treatments at the rate of 1 to 4doses per day, in accordance with the physiological requirements of eachparticular patient. Generally, the active product may be administeredorally 1 to 4 times per day. It goes without saying that, for otherpatients, it will be necessary to prescribe not more than one or twodoses per day.

Another aspect of the present invention relates to a method of treatingcancer, immunologic disorders, autoimmune disorders, neurodegenerativedisorders, or inflammatory disorders in a subject or for providingimmunosuppression for transplanted organs or tissues in a subject. Thismethod includes administering to the subject in need thereof a compoundof the Formula (I):

wherein

L is —(CR³R^(x))_(p)—;

M is —(CR²R^(y))_(r)—;

R¹ is selected from the group consisting of monocyclic and bicyclicaryl, biphenyl, monocyclic and bicyclic heteroaryl, monocyclic andbicyclic heterocyclyl, and monocyclic and bicyclic non-aromaticheterocycle, wherein monocyclic and bicyclic aryl, biphenyl, monocyclicand bicyclic heteroaryl, monocyclic and bicyclic heterocyclyl, andmonocyclic and bicyclic non-aromatic heterocycle can be optionallysubstituted from 1 to 3 times with a substituent selected independentlyat each occurrence thereof from the group consisting of halogen, cyano,—CF₃, C₁₋₆ alkyl, and C₁₋₆ alkoxy;

R² is independently selected at each occurrence thereof from the groupconsisting of H, D, C₁₋₆ alkyl, —CH₂OC₁₋₆ alkyl, —CH₂Ar, and—CH₂heteroaryl, wherein aryl (Ar) can be optionally substituted from 1to 3 times with a substituent selected independently at each occurrencethereof from the group consisting of halogen, cyano, C₁₋₆ alkyl, andC₁₋₆ alkoxy;

R³ is independently selected at each occurrence thereof from the groupconsisting of H, D, —CH₂OC₁₋₆ alkyl, —(CH₂)_(m)C(O)NHR⁵,—(CH₂)_(m)C(O)NR⁶R⁷, —(CH₂)_(m)C(O)OH, and —(CH₂)_(m)C(O)OBn_(;)

R⁴ is selected from the group consisting of H, —C(O)(CH₂)_(n)Ph,—C(O)CH₂NR⁶R⁷, —SO₂Ar, —SO₂C₁₋₆ alkyl, —SO₂C₃₋₆ cycloalkyl,—C(O)(CH₂)_(n)Het, —C(O)C(O)Het, —C(O)C₁₋₆ alkyl, —C(O)OC₁₋₆ alkyl,—C(O)CF₃, heteroaryl, and —(CH₂)_(n)NR⁶R⁷, wherein aryl (Ar) andheteroaryl (Het) can be optionally substituted from 1 to 3 times with asubstituent selected independently at each occurrence thereof from thegroup consisting of halogen, cyano, C₁₋₆ alkyl, and C₁₋₆ alkoxy;

R⁵ is selected from the group consisting of C₁₋₆ alkyl, C₁₋₆ alkoxy,non-aromatic heterocycle, —NR⁶R⁷, and —CR⁸R⁹;

R⁶, R⁷, R⁸, and R⁹ are each independently selected from the groupconsisting of H, D, C₁₋₆ alkyl, and —(CH₂)_(k)OH;

or R⁶ and R⁷ are taken together with the nitrogen to which they areattached to form a morpholine ring;

or R⁸ and R⁹ are taken together with the carbon to which they areattached to form an oxetane ring;

R^(x) is independently selected at each occurrence thereof from thegroup consisting of H, D, —CH₂OC₁₋₆ alkyl, —(CH₂)_(m)C(O)NHR⁵,—(CH₂)_(m)C(O)NR⁶R⁷, and —CH₂C(O)R⁵ _(;)

R^(y) is independently selected at each occurrence thereof from thegroup consisting of H, D, C₁₋₆ alkyl, —CH₂OC₁₋₆ alkyl, —CH₂Ar, and—CH₂heteroaryl, wherein aryl (Ar) can be optionally substituted from 1to 3 times with a substituent selected independently at each occurrencethereof from the group consisting of halogen, cyano, C₁₋₆ alkyl, andC₁₋₆ alkoxy;

X is —(CH₂)_(q)—, —O—, or —(CD₂)_(q)—;

Y is O or S;

k is 1, 2, or 3;

m is 0, 1, 2, 3, 4, or 5;

n is 0, 1, 2, or 3;

p is 1 or 2;

q is 0, 1, or 2;

r is 1 or 2; and

s is 0 or 1;

with a proviso that when s is 0, then r is 2; and when s is 1, thenr+p≥3.

Yet another aspect of the present invention relates to a method oftreating cancer, immunologic disorders, autoimmune disorders,neurodegenerative disorders, or inflammatory disorders in a subject orfor providing immunosuppression for transplanted organs or tissues in asubject. This method includes administering to the subject in needthereof a compound of the Formula (Ia), Formula (Ib), or Formula (Ic):

wherein

R¹ is selected from the group consisting of monocyclic and bicyclicaryl, biphenyl, monocyclic and bicyclic heteroaryl, monocyclic andbicyclic heterocyclyl, and monocyclic and bicyclic non-aromaticheterocycle, wherein monocyclic and bicyclic aryl, biphenyl, monocyclicand bicyclic heteroaryl, monocyclic and bicyclic heterocyclyl, andmonocyclic and bicyclic non-aromatic heterocycle can be optionallysubstituted from 1 to 3 times with a substituent selected independentlyat each occurrence thereof from the group consisting of halogen, cyano,—CF₃, C₁₋₆ alkyl, and C₁₋₆ alkoxy;

R² is independently selected at each occurrence thereof from the groupconsisting of H, D, C₁₋₆ alkyl, —CH₂OC₁₋₆ alkyl, —CH₂Ar, and—CH₂heteroaryl, wherein aryl (Ar) can be optionally substituted from 1to 3 times with a substituent selected independently at each occurrencethereof from the group consisting of halogen, cyano, C₁₋₆ alkyl, andC₁₋₆ alkoxy;

R³ is independently selected at each occurrence thereof from the groupconsisting of H, D, —CH₂OC₁₋₆ alkyl, —(CH₂)_(m)C(O)NHR⁵, and—(CH₂)_(m)C(O)NR⁶R⁷ _(;)

R⁴ is selected from the group consisting of —C(O)(CH₂)_(n)Ph,—C(O)CH₂NR⁶R⁷, —SO₂Ar, —SO₂C₁₋₆ alkyl, —SO₂C₃₋₆ cycloalkyl,—C(O)(CH₂)_(n)Het, —C(O)C(O)Het, —C(O)C₁₋₆ alkyl, —C(O)OC₁₋₆ alkyl,—C(O)CF₃, heteroaryl, and —(CH₂)_(n)NR⁶R⁷, wherein aryl (Ar) andheteroaryl (Het) can be optionally substituted from 1 to 3 times with asubstituent selected independently at each occurrence thereof from thegroup consisting of halogen, cyano, C₁₋₆ alkyl, and C₁₋₆ alkoxy;

R⁵ is selected from the group consisting of C₁₋₆ alkyl, C₁₋₆ alkoxy,non-aromatic heterocycle, —NR⁶R⁷, and —CR⁸R⁹;

R⁶, R⁷, R⁸, and R⁹ are each independently selected from the groupconsisting of H, D, C₁₋₆ alkyl, and —(CH₂)_(k)OH;

or R⁶ and R⁷ are taken together with the nitrogen to which they areattached to form a morpholine ring;

or R⁸ and R⁹ are taken together with the carbon to which they areattached to form an oxetane ring;

R^(x) is independently selected at each occurrence thereof from thegroup consisting of H, D, —CH₂OC₁₋₆ alkyl, —(CH₂)_(m)C(O)NHR⁵,—(CH₂)_(m)C(O)NR⁶R⁷, and —CH₂C(O)R⁵ _(;)

R^(y) is independently selected at each occurrence thereof from thegroup consisting of H, D, C₁₋₆ alkyl, —CH₂OC₁₋₆ alkyl, —CH₂Ar, and—CH₂heteroaryl, wherein aryl (Ar) can be optionally substituted from 1to 3 times with a substituent selected independently at each occurrencethereof from the group consisting of halogen, cyano, C₁₋₆ alkyl, andC₁₋₆ alkoxy;

X is —(CH₂)_(q)—, —O—, or —(CD₂)_(q)—;

Y is O or S;

k is 1, 2, or 3;

m is 0, 1, 2, 3, 4, or 5;

n is 0, 1, 2, or 3;

q is 0, 1, or 2; and

s is 0 or 1.

In one embodiment, an autoimmune disorder is treated. The autoimmunedisorder is selected from the group consisting of arthritis, colitis,multiple sclerosis, lupus, systemic sclerosis, and sjögren syndrome.Alternatively, the autoimmune disorder is selected from the groupconsisting of arthritis, colitis, multiple sclerosis, and lupus.

In another embodiment, immunosuppression is provided for transplantedorgans or tissues. The immunosuppression is used to prevent transplantrejection and graft-verse-host disease.

In another embodiment, an inflammatory disorder is treated. Theinflammatory disorder is Crohn's disease, and ulcerative colitis.Alternatively, the inflammatory disorder is Crohn's disease.

In yet another embodiment, cancer is treated. The cancer is selectedfrom the group consisting of neoplastic disorders, hematologicmalignancies, and lymphocytic malignancies.

EXAMPLES

The following examples are provided to illustrate embodiments of thepresent invention but are by no means intended to limit its scope.

Example 1 General Procedure for HATU Coupling

Carboxylic acid (1.0 eq.),O-(7-Azabenzotriazol-1-yl)-N,N,N,N′-tetramethyluroniumhexafluorophosphate (HATU) (1.2 eq.), and 1-Hydroxy-7-Azabenzotriazole(HOAt) 0.6M in DMF (1.0 eq.) were dissolved in DMF under argonatmosphere. The solution was cooled to 0° C. and amine was added. Afterstirring for 5 minutes at 0° C., Hünig's base (3-4 eq.) was added. Thereaction mixture was stirred at 0° C. for 1 hour. After completion ofreaction (1 hour, monitored by LCMS), water (10 mL) was added toreaction mixture and stirred 30 minutes. Product was isolated either byethyl acetate extraction or filtering the precipitate.

Example 2 General Procedure for EDC Coupling

Carboxylic acid (1.0 eq.),N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC) (1.2eq.), and 1-Hydroxybenzotriazole (HOBt) (1.3 eq.) were dissolved in DMFunder argon atmosphere. The solution was cooled to 0° C. andtert-butylamine was added. After stirring for 5 minutes at 0° C.,Hünig's base (2-3 eq.) was added. The reaction mixture was allowed towarm to room temperature slowly and stirred at room temperatureovernight.

Example 3 General Procedure for Boc-Deprotection

The substrate was dissolved in dichloromethane and the solution wascooled to 0° C. Trifluoroacetic acid (20% v/v with respect todichloromethane) was added to the solution drop wise at 0° C. withconstant stirring. The mixture was allowed to warm to room temperatureslowly (over a period of 1 hour), and stirred until the completion ofreaction (monitored by LCMS). Excess trifluoroacetic acid anddichloromethane were evaporated and crude was dried under vacuum.

Example 4 General Procedure for O-Debenzylation

The substrate was dissolved in methanol. Palladium on carbon (10%) wasadded carefully. Residual air from the flask was removed and the mixturewas stirred at room temperature for 3-4 hours under hydrogen atmosphereusing a hydrogen balloon. After completion of reaction, the mixture wasfiltered through celite. Filtrate was evaporated and dried under vacuumto give product.

Example 5 General Procedure for N-Sulfonamide Preparation of Amines

The primary amine (generally TFA salt) was dissolved in dichloromethane.The solution was cooled to 0° C. and triethylamine (2.0-3.0 eq.) wasadded. Sulfonyl chloride (1.5 eq.) was added to the solution in oneportion and reaction mixture was warmed to room temperature (over 15minutes). After completion of reaction (2-3 hours), dichloromethane wasevaporated and crude was purified by HPLC to give pure product.

Example 6 Preparation of tert-butyl(S)-(4-((naphthalen-1-ylmethyl)amino)-4-oxobutan-2-yl)carbamate(PKS2241, PKS2261)

The title compound was synthesized by following the general protocol forHATU mediated coupling of Boc-L-β-homoalanine and 1-naphthylmethylamineon a 1.5 mmol scale. After completion of reaction, water was added toreaction mixture to give white precipitate. Precipitate was filtered,washed with water and dried in air to give product (490 mg, 95%) as awhite solid. ¹H NMR (500 MHz, DMSO-d₆) δ 8.38 (t, J=5.7 Hz, 1H),8.06-8.04 (m, 1H), 7.95-7.93 (m, 1H), 7.86-7.84 (m, 1H), 7.56-7.52 (m,2H), 7.48-7.42 (m, 2H), 6.72 (d, J=8.3 Hz, 1H), 4.76-4.68 (m, 2H),3.89-3.80 (m, 1H), 2.35 (dd, J=13.9, 5.7 Hz, 1H), 2.18 (dd, J=13.9, 8.2Hz, 1H), 1.37 (s, 9H), 1.01 (d, J=6.5 Hz, 3H).

Example 7 Preparation of (S)-3-amino-N-(naphthalen-1-ylmethyl)butanamide2,2,2-trifluoroacetate (PKS2245, PKS2262)

The title compound was prepared by following the general protocol forBoc-deprotection of PKS2261 (480 mg, 1.40 mmol). The crude yellow pastewas triturated with diethyl ether and kept standing overnight. The whitesolid was filtered and dried to give product (480 mg, 96%). ¹H NMR (500MHz, DMSO-d₆) δ 8.69 (t, J=5.7 Hz, 1H), 8.06-8.05 (m, 1H), 7.97-7.95 (m,1H), 7.88 (dd, J=7.4, 2.0 Hz, 1H), 7.80 (bs, 3H), 7.58-7.53 (m, 2H),7.50-7.45 (m, 2H), 4.79 (dd, J=15.0, 5.7 Hz, 1H), 4.73 (dd, J=15.0, 5.5Hz, 1H), 3.57-3.50 (m, 1H), 2.51-2.43 (m, 2H), 1.17 (d, J=6.5 Hz, 3H).

Example 8 Preparation of benzyl(S)-3-((tert-butoxycarbonyl)amino)-4-(((S)-4-((naphthalen-1-ylmethyl)amino)-4-oxobutan-2-yl)amino)-4-oxobutanoate(PKS2265)

The title compound was synthesized by following the general protocol forHATU mediated coupling of Boc-Asp(OBn)-OH (142.3 mg, 0.44 mg) andPKS2262 (143 mg, 0.40 mmol). After completion of reaction, water wasadded to reaction mixture to give white precipitate. Precipitate wasfiltered, washed with water and dried in air to give product (202 mg,92%) as a white solid. ¹H NMR (500 MHz, DMSO-d₆) δ 8.41 (t, J=5.8 Hz,1H), 8.06-8.04 (m, 1H), 7.95-7.93 (m, 1H), 7.86 (d, J=8.3 Hz, 1H), 7.84(d, J=7.7 Hz, 1H), 7.56-7.51 (m, 2H), 7.47-7.42 (m, 2H), 7.36-7.29 (m,5H), 7.11 (d, J=8.3 Hz, 1H), 5.10-5.05 (m, 2H), 4.76-4.68 (m, 2H),4.30-4.25 (m, 1H), 4.14-4.08 (m, 1H), 2.73-2.69 (m, 1H), 2.57 (dd,J=16.1, 8.8 Hz, 1H), 2.34 (dd, J=14.1, 5.6 Hz, 1H), 2.23 (dd, J=14.1,7.5 Hz, 1H), 1.37 (s, 9H), 1.03 (d, J=6.6 Hz, 3H).

Example 9 Preparation of(S)-3-((tert-butoxycarbonyl)amino)-4-(((S)-4-((naphthalen-1-ylmethyl)amino)-4-oxobutan-2-yl)amino)-4-oxobutanoicacid (PKS2267)

The title compound was synthesized by following the O-debenzylationprotocol of PKS2265 (202 mg, 0.37 mmol). Product (168 mg, quant.) wasisolated as a white solid. ¹H NMR (500 MHz, DMSO-d₆) δ 8.44-8.41 (m,1H), 8.06-8.04 (m, 1H), 7.95-7.94 (m, 1H), 7.85 (d, J=8.0 Hz, 1H), 7.81(d, J=8.3 Hz, 1H), 7.57-7.52 (m, 2H), 7.48-7.43 (m, 2H), 7.02 (d, J=8.2Hz, 1H), 4.75 (dd, J=15.1, 5.7 Hz, 1H), 4.70 (dd, J=15.1, 5.7 Hz, 1H),4.21-4.16 (m, 1H), 4.13-4.08 (m, 1H), 2.58 (dd, J=16.4, 5.1 Hz, 1H),2.43 (dd, J=16.4, 8.5 Hz, 1H), 2.35 (dd, J=14.1, 5.4 Hz, 1H), 2.23 (dd,J=14.1, 7.7 Hz, 1H), 1.38 (s, 9H), 1.03 (d, J=6.6 Hz, 3H).

Example 10 Preparation tert-butyl((S)-4-(tert-butylamino)-1-(((S)-4-((naphthalen-1-ylmethyl)amino)-4-oxobutan-2-yl)amino)-1,4-dioxobutan-2-yl)carbamate(PKS2247, PKS2272)

The title compound was synthesized following the general protocol of EDCmediated coupling of PKS2267 (168 mg, 0.367 mmol) and tert-butyl amine(58 μL, 0.551 mmol). A white precipitate appeared which was filtered,washed with water and dried in air to give product (170 mg, 90%) as awhite solid. ¹H NMR (500 MHz, DMSO-d₆) δ 8.43 (t, J=5.6 Hz, 1H),8.06-8.04 (m, 1H), 7.95-7.94 (m, 1H), 7.86-7.81 (m, 2H), 7.57-7.52 (m,2H), 7.48-7.43 (m, 2H), 7.34 (s, 1H), 6.76 (d, J=8.3 Hz, 1H), 4.75 (dd,J=15.1, 5.7 Hz, 1H), 4.70 (dd, J=15.1, 5.5 Hz, 1H), 4.19-4.07 (m, 2H),2.36-2.21 (m, 4H), 1.37 (s, 9H), 1.22 (s, 9H), 1.03 (d, J=6.6 Hz, 3H).

Example 11 Preparation of(S)-2-amino-N⁴-(tert-butyl)-N¹-((S)-4-((naphthalen-1-ylmethyl)amino)-4-oxobutan-2-yl)succinamide2,2,2-trifluoroacetate (PKS2248)

The title compound was prepared by following the general protocol forBoc-Deprotection of PKS2272 (62 mg, 0.12 mmol). The crude was treatedwith diethyl ether and kept standing 3 hours. The white solid wasfiltered and dried to give product (60 mg, 95%). ¹H NMR (500 MHz,DMSO-d₆) δ 8.47 (t, J=5.6 Hz, 1H), 8.32 (d, J=7.9 Hz, 1H), 8.06-8.03 (m,4H), 7.96-7.95 (m, 1H), 7.87-7.85 (m, 1H), 7.80 (s, 1H), 7.57-7.53 (m,2H), 7.49-7.43 (m, 2H), 4.78 (dd, J=15.1, 5.8 Hz, 1H), 4.69 (dd, J=15.1,5.4 Hz, 1H), 4.21-4.13 (m, 1H), 3.95 (m, 1H), 2.60 (dd, J=16.7, 4.6 Hz,1H), 2.55-2.49 (m, 1H), 2.34 (dd, J=14.1, 5.7 Hz, 1H), 2.26 (dd, J=14.1,8.0 Hz, 1H), 1.26 (s, 9H), 1.07 (d, J=6.6 Hz, 3H).

Example 12 Preparation of(S)-N⁴-(tert-butyl)-2-(5-methylisoxazole-3-carboxamido)-N¹-((S)-4-((naphthalen-1-ylmethyl)amino)-4-oxobutan-2-yl)succinamide(PKS2249)

The title compound was prepared by following the general protocol forHATU mediated coupling of 5-methylisoxazole-3-carboxylic acid (5.6 mg,0.044 mmol) and PKS2248 (21.1 mg, 0.04 mmol). After completion ofreaction, the mixture was purified by HPLC to give pure product (14.4mg, 69%) as a white solid. ¹H NMR (500 MHz, DMSO-d₆) δ 8.50 (d, J=8.1Hz, 1H), 8.41 (t, J=5.7 Hz, 1H), 8.05-8.03 (m, 1H), 7.99 (d, J=8.1 Hz,1H), 7.95-7.93 (m, 1H), 7.84 (d, J=8.0 Hz, 1H), 7.57-7.51 (m, 2H),7.48-7.42 (m, 3H), 6.53 (s, 1H), 4.74 (dd, J=15.1, 5.7 Hz, 1H), 4.68(dd, J=15.1, 5.5 Hz, 1H), 4.63 (td, J=8.4, 4.8 Hz, 1H), 4.16-4.08 (m,1H), 2.54 (dd, J=14.3, 8.7 Hz, 1H), 2.45 (s, 3H), 2.41 (dd, J=14.3, 4.8Hz, 1H), 2.35 (dd, J=14.0, 5.7 Hz, 1H), 2.25 (dd, J=14.0, 7.7 Hz, 1H),1.18 (s, 9H), 1.04 (d, J=6.6 Hz, 3H). ¹³C NMR (126 MHz, DMSO) δ 171.34,169.90, 169.22, 168.86, 158.58, 158.24, 134.57, 133.32, 130.89, 128.53,127.62, 126.25, 125.85, 125.62, 125.43, 123.52, 101.35, 50.51, 50.14,42.57, 41.61, 40.19, 38.33, 28.38, 19.98, 11.86.

Example 13 Preparation of(S)-N⁴-(tert-butyl)-N¹-((S)-4-((naphthalen-1-ylmethyl)amino)-4-oxobutan-2-yl)-2-(3-phenylpropanamido)succinamide(PKS2251)

The title compound was prepared by following the general protocol forHATU mediated coupling of 3-phenylpropanoic acid (5.0 mg, 0.033 mmol)and PKS2248 (15.8 mg, 0.03 mmol). After completion of reaction, themixture was purified by HPLC to give pure product (14.3 mg, 88%) as awhite solid. ¹H NMR (500 MHz, DMSO-d₆) δ 8.44 (t, J=5.7 Hz, 1H),8.06-8.04 (m, 1H), 7.99 (d, J=8.1 Hz, 1H), 7.95-7.93 (m, 1H), 7.86-7.83(m, 2H), 7.57-7.53 (m, 2H), 7.49-7.43 (m, 2H), 7.33 (s, 1H), 7.26-7.23(m, 2H), 7.20-7.14 (m, 3H), 4.75 (dd, J=15.1, 5.7 Hz, 1H), 4.71 (dd,J=15.1, 5.6 Hz, 1H), 4.48 (td, J=8.2, 5.6 Hz, 1H), 4.14-4.06 (m, 1H),2.82-2.78 (m, 2H), 2.44-2.33 (m, 4H), 2.29 (dd, J=14.6, 8.3 Hz, 1H),2.23 (dd, J=14.1, 7.8 Hz, 1H), 1.21 (s, 9H), 1.03 (d, J=6.6 Hz, 3H). ¹³CNMR (126 MHz, DMSO) δ 171.10, 169.97, 169.92, 168.75, 141.31, 134.56,133.28, 130.86, 128.49, 128.25, 128.11, 127.57, 126.19, 125.82, 125.79,125.56, 125.38, 123.48, 50.10, 50.02, 42.32, 41.50, 40.15, 38.73, 36.89,31.00, 28.44, 19.87.

Example 14 Preparation of(S)-N⁴-(tert-butyl)-2-((4-methylphenyl)sulfonamido)-N¹-((S)-4-((naphthalen-1-ylmethyl)amino)-4-oxobutan-2-yl)succinamide(PKS2252)

The title compound was prepared by following the general procedure forN-sulfonamide formation of PKS2248 (21.1 mg, 0.04 mmol) with tosylchloride (11.5 mg, 0.06 mmol). The product was isolated as white solid(12.7 mg, 56%) after HPLC purification. ¹H NMR (500 MHz, DMSO-d₆) δ 8.38(t, J=5.7 Hz, 1H), 8.05-8.03 (m, 1H), 7.95-7.93 (m, 1H), 7.84 (d, J=8.1Hz, 1H), 7.78 (d, J=8.1 Hz, 1H), 7.71 (bs, 1H), 7.65 (d, J=8.2 Hz, 2H),7.56-7.52 (m, 2H), 7.48-7.41 (m, 2H), 7.32-7.29 (m, 3H), 4.73 (dd,J=15.1, 5.7 Hz, 1H), 4.68 (dd, J=15.1, 5.6 Hz, 1H), 4.01 (m, 1H),3.94-3.85 (m, 1H), 2.33 (s, 3H), 2.29-2.15 (m, 3H), 2.11 (dd, J=14.1,8.2 Hz, 1H), 1.18 (s, 9H), 0.83 (d, J=6.6 Hz, 3H). ¹³C NMR (126 MHz,DMSO) δ 169.67, 168.87, 168.02, 142.36, 138.33, 134.59, 133.28, 130.86,129.24, 128.48, 127.57, 126.63, 126.17, 125.79, 125.60, 125.37, 123.50,53.60, 50.07, 42.18, 41.52, 40.13, 39.39, 28.40, 20.91, 19.43.

Example 15 Preparation of(S)-N⁵-(tert-butyl)-N¹-((S)-4-((naphthalen-1-ylmethyl)amino)-4-oxobutan-2-yl)-2-(3-phenylpropanamido)pentanediamide(PKS2253)

The title compound was synthesized by following the general protocol forHATU mediated coupling of PhCH₂CH₂C(O)-Glu(NHtBu)-OH (18.4 mg, 0.055mmol) and H-β-homo-Ala-CH₂-naphth TFA salt (17.8 mg, 0.05 mmol).Purification by HPLC provided the product (23.5 mg, 84%) as a whitesolid. ¹H NMR (500 MHz, DMSO-d₆) δ 8.45 (t, J=5.7 Hz, 1H), 8.07-8.05 (m,1H), 8.00-7.93 (m, 2H), 7.89 (d, J=8.1 Hz, 1H), 7.85-7.83 (m, 1H),7.57-7.51 (m, 2H), 7.48-7.43 (m, 2H), 7.34 (s, 1H), 7.27-7.23 (m, 2H),7.21-7.14 (m, 3H), 4.77 (dd, J=15.1, 5.7 Hz, 1H), 4.70 (dd, J=15.1, 5.6Hz, 1H), 4.18-4.13 (m, 2H), 2.81 (t, J=7.9 Hz, 2H), 2.46-2.42 (m, 2H),2.38 (dd, J=14.0, 5.6 Hz, 1H), 2.24 (dd, J=14.0, 7.8 Hz, 1H), 1.99 (t,J=8.1 Hz, 2H), 1.82-1.75 (m, 1H), 1.70-1.62 (m, 1H), 1.21 (s, 9H), 1.05(d, J=6.6 Hz, 3H). ¹³C NMR (126 MHz, DMSO) δ 171.24, 171.08, 170.30,169.92, 141.35, 134.54, 133.28, 130.85, 128.48, 128.22, 128.16, 127.56,126.19, 125.80, 125.52, 125.39, 123.49, 52.30, 49.80, 42.27, 41.82,40.14, 36.78, 32.57, 31.06, 28.54, 28.51, 20.12.

Example 16 Preparation of(S)-N⁴-(tert-butyl)-N¹-((S)-4-((naphthalen-1-ylmethyl)amino)-4-oxobutan-2-yl)-2-(phenylsulfonamido)succinamide(PKS2260)

The title compound was prepared by following the general procedure forN-sulfonamide formation of PKS2248 (12.1 mg, 0.023 mmol) withphenylsulfonyl chloride (4 μL, 0.028 mmol). The product was isolated asa white solid (10.0 mg, 79%) after HPLC purification. ¹H NMR (500 MHz,DMSO-d₆) δ 8.37 (t, J=5.7 Hz, 1H), 8.04-8.02 (m, 1H), 7.95-7.93 (m, 1H),7.85-7.81 (m, 3H), 7.77 (d, J=7.6 Hz, 2H), 7.60-7.41 (m, 7H), 7.32 (s,1H), 4.73 (dd, J=15.1, 5.8 Hz, 1H), 4.68 (dd, J=15.1, 5.5 Hz, 1H),4.07-4.03 (m, 1H), 3.92-3.86 (m, 1H), 2.28-2.17 (m, 3H), 2.12 (dd,J=14.1, 8.3 Hz, 1H), 1.18 (s, 9H), 0.83 (d, J=6.6 Hz, 3H). ¹³C NMR (126MHz, DMSO-d₆) δ 169.67, 168.85, 167.98, 141.25, 134.59, 133.28, 132.17,130.86, 128.80, 128.48, 127.57, 126.54, 126.19, 125.80, 125.60, 125.38,123.50, 53.62, 50.07, 42.16, 41.55, 40.13, 39.69, 28.42, 19.49.

Example 17 Preparation of(S)-N⁴-(tert-butyl)-2-(cyclopropanesulfonamido)-N¹-((S)-4-((naphthalen-1-ylmethyl)amino)-4-oxobutan-2-yl)succinamide(PKS2295)

The title compound was prepared by following the general procedure forN-sulfonamide formation of PKS2248 (21.1 mg, 0.04 mmol) withcyclopropylsulfonyl chloride (6 μL, 0.06 mmol). The product was isolatedas a white solid (16.3 mg, 79%) after HPLC purification. ¹H NMR (500MHz, DMSO-d₆) δ 8.42 (t, J=5.6 Hz, 1H), 8.06-8.04 (m, 1H), 7.97-7.94 (m,2H), 7.85 (d, J=7.9 Hz, 1H), 7.57-7.52 (m, 2H), 7.48-7.43 (m, 2H), 7.40(s, 1H), 7.26 (d, J=9.1 Hz, 1H), 4.75 (dd, J=15.1, 5.7 Hz, 1H), 4.70(dd, J=15.1, 5.5 Hz, 1H), 4.17-4.06 (m, 2H), 2.50-2.46 (m, 1H),2.41-2.32 (m, 3H), 2.25 (dd, J=14.1, 8.0 Hz, 1H), 1.23 (s, 9H), 1.04 (d,J=6.6 Hz, 3H), 0.90-0.82 (m, 4H). ¹³C NMR (126 MHz, DMSO-d₆) δ 169.93,169.75, 168.37, 134.57, 133.28, 130.87, 128.48, 127.58, 126.19, 125.80,125.62, 125.38, 123.51, 53.71, 50.12, 42.41, 41.59, 40.15, 39.82, 30.28,28.45, 19.80, 5.13, 4.84.

Example 18 Preparation of benzyl(R)-3-((tert-butoxycarbonyl)amino)-5-(((S)-1-((naphthalen-1-ylmethyl)amino)-1-oxopropan-2-yl)amino)-5-oxopentanoate(PKS2266)

The title compound was synthesized by following the general protocol forHATU mediated coupling of N-Boc-L-β-glutamic acid 5-benzyl ester (84 mg,0.25 mmol) and H-Ala-CH₂-naphth TFA salt (94 mg, 0.275 mmol). Aftercompletion of reaction, water was added to reaction mixture to givewhite precipitate. Precipitate was filtered, washed with water and driedin air to give product (115 mg, 84%) as a white solid. ¹H NMR (500 MHz,DMSO-d₆) δ 8.40 (t, J=5.6 Hz, 1H), 8.11 (d, J=7.4 Hz, 1H), 8.04-8.02 (m,1H), 7.95-7.93 (m, 1H), 7.83 (d, J=7.9 Hz, 1H), 7.56-7.51 (m, 2H),7.47-7.41 (m, 2H), 7.37-7.30 (m, 5H), 6.82 (d, J=8.3 Hz, 1H), 5.05 (s,2H), 4.77-4.69 (m, 2H), 4.33-4.28 (m, 1H), 4.22-4.13 (m, 1H), 2.55 (dd,J=15.0, 5.0 Hz, 1H), 2.49-2.44 (m, 1H), 2.38 (dd, J=14.6, 6.3 Hz, 1H),2.32 (dd, J=14.6, 7.5 Hz, 1H), 1.34 (s, 9H), 1.21 (d, J=7.1 Hz, 3H).

Example 19 Preparation of(R)-3-((tert-butoxycarbonyl)amino)-5-(((S)-1-((naphthalen-1-ylmethyl)amino)-1-oxopropan-2-yl)amino)-5-oxopentanoicacid (PKS2268)

The title compound was synthesized by following the O-debenzylationprotocol of PKS2266 (110 mg, 0.2 mmol). Product (92 mg, quant.) wasisolated as a white solid. ¹H NMR (500 MHz, DMSO-d₆) δ 8.49 (t, J=5.8Hz, 1H), 8.16 (d, J=7.4 Hz, 1H), 8.04 (d, J=7.8 Hz, 1H), 7.94 (d, J=7.3Hz, 1H), 7.83 (d, J=7.8 Hz, 1H), 7.56-7.51 (m, 2H), 7.48-7.42 (m, 2H),6.74 (d, J=8.2 Hz, 1H), 4.74-4.71 (m, 2H), 4.33-4.28 (m, 1H), 4.11-4.04(m, 1H), 2.41-2.29 (m, 4H), 1.35 (s, 9H), 1.23 (d, J=7.1 Hz, 3H).

Example 20 Preparation of tert-butyl((R)-1-(tert-butylamino)-5-(((S)-1-((naphthalen-1-ylmethyl)amino)-1-oxopropan-2-yl)amino)-1,5-dioxopentan-3-yl)carbamate(PKS2271)

The title compound was prepared by following the general protocol forEDC mediated coupling of PKS2268 (92 mg, 0.2 mmol) and tert-butylamine(31.5 μL, 0.3 mmol). After completion of reaction, water was added toreaction mixture to give white precipitate. Precipitate was filtered,washed with water and dried in air to give product (85 mg, 83%) as awhite solid. ¹H NMR (500 MHz, DMSO-d₆) δ 8.44 (t, J=5.9 Hz, 1H),8.05-8.03 (m, 2H), 7.95-7.93 (m, 1H), 7.84 (d, J=7.7 Hz, 1H), 7.56-7.53(m, 2H), 7.48-7.43 (m, 2H), 7.34 (s, 1H), 6.56 (d, J=8.4 Hz, 1H),4.78-4.69 (m, 2H), 4.34-4.28 (m, 1H), 4.10-4.00 (m, 1H), 2.36-2.12 (m,4H), 1.35 (s, 9H), 1.23-1.22 (m, 12H).

Example 21 Preparation of(R)-3-amino-N¹-(tert-butyl)-N⁵-((S)-1-((naphthalen-1-ylmethyl)amino)-1-oxopropan-2-yl)pentanediamide2,2,2-trifluoroacetate (PKS2273)

The title compound was synthesized by following the general protocol forBoc-deprotection of PKS2271 (80 mg, 0.156 mmol). ¹H NMR (500 MHz,DMSO-d₆) δ 8.54 (t, J=5.7 Hz, 1H), 8.46 (d, J=7.3 Hz, 1H), 8.05-8.03 (m,1H), 7.96-7.94 (m, 1H), 7.89-7.82 (m, 5H), 7.56-7.54 (m, 2H), 7.49-7.43(m, 2H), 4.78 (dd, J=15.4, 5.9 Hz, 1H), 4.70 (dd, J=15.4, 5.5 Hz, 1H),4.39-4.33 (m, 1H), 3.67-3.61 (m, 1H), 2.53-2.49 (m, 2H), 2.42 (dd,J=15.9, 5.8 Hz, 1H), 2.36 (dd, J=15.9, 7.2 Hz, 1H), 1.26-1.24 (m, 12H).

Example 22 Preparation of(R)-N¹-(tert-butyl)-N⁵-((S)-1-((naphthalen-1-ylmethyl)amino)-1-oxopropan-2-yl)-3-(3-phenylpropanamido)pentanediamide(PKS2278)

The title compound was synthesized by following the general protocol forHATU mediated coupling of 3-phenylpropanoic acid (6.6 mg, 0.044 mmol)and PKS2273 (21.1 mg, 0.04 mmol). After completion of reaction, mixturewas purified by HPLC to give pure product (20.7 mg, 95%) as a whitesolid. ¹H NMR (500 MHz, DMSO-d₆) δ 8.44-8.42 (m, 1H), 8.07 (d, J=7.3 Hz,1H), 8.02-8.00 (m, 1H), 7.95-7.93 (m, 1H), 7.83 (d, J=7.9 Hz, 1H), 7.68(d, J=8.1 Hz, 1H), 7.54-7.52 (m, 2H), 7.45-7.39 (m, 2H), 7.37 (s, 1H),7.24-7.21 (m, 2H), 7.16-7.13 (m, 3H), 4.71 (d, J=5.7 Hz, 2H), 4.33-4.29(m, 2H), 2.76-2.73 (m, 2H), 2.37-2.16 (m, 6H), 1.24-1.22 (m, 12H). ¹³CNMR (126 MHz, DMSO) δ 172.36, 170.51, 169.71, 169.42, 141.31, 134.41,133.21, 130.74, 128.46, 128.23, 128.07, 127.39, 126.12, 125.79, 125.73,125.37, 124.90, 123.32, 49.96, 48.42, 44.21, 40.73, 40.11, 39.75, 37.21,31.14, 28.45, 18.27.

Example 23 Preparation of(R)-N¹-(tert-butyl)-3-(4-methylphenylsulfonamido)-N⁵-((S)-1-((naphthalen-1-ylmethyl)amino)-1-oxopropan-2-yl)pentanediamide(PKS2279)

The title compound was prepared by following the general procedure forN-sulfonamide formation of PKS2273 (21.1 mg, 0.04 mmol) with tosylchloride (11.4 mg, 0.06 mmol). The product was isolated as a white solid(17.8 mg, 78%) after HPLC purification. ¹H NMR (500 MHz, DMSO-d₆) δ 8.42(t, J=5.8 Hz, 1H), 8.08-8.03 (m, 2H), 7.95-7.94 (m, 1H), 7.84 (d, J=7.9Hz, 1H), 7.66 (d, J=8.1 Hz, 2H), 7.55-7.53 (m, 2H), 7.48-7.42 (m, 3H),7.39 (s, 1H), 7.32 (d, J=8.1 Hz, 2H), 4.78-4.70 (m, 2H), 4.28-4.23 (m,1H), 3.81-3.74 (m, 1H), 2.35 (s, 3H), 2.29-2.20 (m, 2H), 2.13 (d, J=6.6Hz, 2H), 1.20-1.18 (m, 12H). ¹³C NMR (126 MHz, DMSO) δ 172.19, 169.22,168.94, 142.43, 138.54, 134.41, 133.24, 130.77, 129.45, 128.48, 127.46,126.50, 126.14, 125.77, 125.38, 125.07, 123.37, 50.05, 48.51, 48.30,40.71, 40.13, 39.95, 28.39, 20.94, 18.26.

Example 24 Preparation of (S)-benzyl4-((tert-butoxycarbonyl)amino)-6-(((S)-1-((naphthalen-1-ylmethyl)amino)-1-oxopropan-2-yl)amino)-6-oxohexanoate(PKS2274)

The title compound was synthesized by following the general protocol forHATU mediated coupling of (S)-3-(Boc-amino)adipic acid 6-benzyl ester(23.7 mg, 0.0675 mmol) and H-Ala-CH₂-naphth TFA salt (23.1 mg, 0.0675mmol). After completion of reaction, mixture was purified by HPLC togive product (23.0 mg, 61%) as a white solid. ¹H NMR (500 MHz,Chloroform-d) δ 7.96 (d, J=8.3 Hz, 1H), 7.86-7.84 (m, 1H), 7.78 (dd,J=7.3, 2.1 Hz, 1H), 7.55-7.47 (m, 2H), 7.42-7.38 (m, 2H), 7.36-7.28 (m,5H), 6.66-6.64 (m, 1H), 6.24 (d, J=7.2 Hz, 1H), 5.13 (d, J=8.9 Hz, 1H),5.05-5.00 (m, 2H), 4.91 (dd, J=14.6, 5.6 Hz, 1H), 4.84 (dd, J=14.6, 5.3Hz, 1H), 4.45-4.39 (m, 1H), 3.75-3.70 (m, 1H), 2.37-2.23 (m, 4H),1.73-1.61 (m, 2H), 1.39 (s, 9H), 1.37 (d, J=7.0 Hz, 3H).

Example 25 Preparation of(S)-4-((tert-butoxycarbonyl)amino)-6-(((S)-1-((naphthalen-1-ylmethyl)amino)-1-oxopropan-2-yl)amino)-6-oxohexanoicacid (PKS2277)

The title compound was synthesized by following the O-debenzylationprotocol of PKS2274 (23 mg, 0.04 mmol). The crude was purified by HPLCto give product (17.5 mg, 91%) as a white solid. ¹H NMR (500 MHz,DMSO-d₆) δ 8.44 (t, J=5.8 Hz, 1H), 8.13-8.11 (m, 1H), 8.04 (d, J=7.9 Hz,1H), 7.95-7.93 (m, 1H), 7.84 (d, J=7.8 Hz, 1H), 7.56-7.51 (m, 2H),7.48-7.42 (m, 2H), 6.71 (d, J=8.8 Hz, 1H), 4.72-4.71 (m, 2H), 4.33-4.31(m, 1H), 3.75 (m, 1H), 2.32-2.14 (m, 4H), 1.70-1.65 (m, 1H), 1.57-1.51(m, 1H), 1.36 (s, 9H), 1.22 (d, J=7.2 Hz, 3H).

Example 26 Preparation of tert-butyl((S)-6-(tert-butylamino)-1-(((S)-1-((naphthalen-1-ylmethyl)amino)-1-oxopropan-2-yl)amino)-1,6-dioxohexan-3-yl)carbamate(PKS2282)

Title compound was prepared by following the general protocol for EDCmediated coupling of PKS2277 (17.5 mg, 0.037 mmol) and tert-butylamine(6.0 μL, 0.056 mmol). After completion of reaction, mixture was purifiedby HPLC to give product (6.8 mg, 35%) as a white solid. ¹H NMR (500 MHz,DMSO-d₆) δ 8.42 (t, J=5.8 Hz, 1H), 8.07-8.03 (m, 2H), 7.95-7.93 (m, 1H),7.84 (d, J=7.9 Hz, 1H), 7.56-7.53 (m, 2H), 7.48-7.42 (m, 2H), 7.33 (s,1H), 6.61 (d, J=8.7 Hz, 1H), 4.78-4.69 (m, 2H), 4.35-4.29 (m, 1H),3.75-3.68 (m, 1H), 2.31-2.22 (m, 2H), 2.02-1.97 (m, 2H), 1.62-1.57 (m,1H), 1.53-1.48 (m, 1H), 1.36 (s, 9H), 1.23-1.21 (m, 12H).

Example 27 Preparation of(S)-3-amino-N⁶-(tert-butyl)-N¹-((S)-1-((naphthalen-1-ylmethyl)amino)-1-oxopropan-2-yl)hexanediamide2,2,2-trifluoroacetate (PKS2289)

The title compound was synthesized by following the general protocol forBoc-deprotection of PKS2282 (6.8 mg, 0.013 mmol). The product was usedin next step without further purification. ¹H NMR (500 MHz,Chloroform-d) δ 8.24 (s, 3H), 8.10 (m, 1H), 7.88-7.80 (m, 3H), 7.55-7.49(m, 2H), 7.42-7.40 (m, 2H), 6.82 (m, 1H), 5.98 (s, 1H), 5.00-4.95 (m,1H), 4.76-4.72 (m, 1H), 4.37 (m, 1H), 3.55 (m, 1H), 2.75-2.72 (m, 1H),2.61-2.57 (m, 1H), 2.40 (t, J=5.4 Hz, 2H), 1.93 (m, 1H), 1.80 (m,1H),1.39-1.38 (m, 3H), 1.32 (s, 9H).

Example 28 Preparation of(S)-N⁶-(tert-butyl)-3-(4-methylphenylsulfonamido)-N¹-((S)-1-((naphthalen-1-ylmethyl)amino)-1-oxopropan-2-yl)hexanediamide(PKS2290)

The title compound was prepared by following the general procedure forN-sulfonamide formation of PKS2289 (crude from previous step, 0.013mmol) with tosyl chloride (5.0 mg, 0.026 mmol). The product was isolatedas a white solid (6.0 mg, 79% for 2 steps) after HPLC purification. ¹HNMR (500 MHz, DMSO-d₆) δ 8.41 (t, J=5.8 Hz, 1H), 8.10 (d, J=7.5 Hz, 1H),8.03-8.01 (m, 1H), 7.96-7.94 (m, 1H), 7.84 (d, J=8.0 Hz, 1H), 7.66 (d,J=8.2 Hz, 2H), 7.59 (d, J=7.8 Hz, 1H), 7.55-7.52 (m, 2H), 7.47-7.40 (m,2H), 7.35 (d, J=8.0 Hz, 2H), 7.31 (s, 1H), 4.74 (dd, J=15.4, 5.8 Hz,1H), 4.70 (dd, J=15.4, 5.7 Hz, 1H), 4.28-4.22 (m, 1H), 3.45-3.39 (m,1H), 2.36 (s, 3H), 2.20-2.00 (m, 3H), 1.94-1.88 (m, 1H), 1.57-1.50 (m,1H), 1.47-1.39 (m, 1H), 1.20 (s, 9H), 1.18 (d, J=7.4 Hz, 4H). ¹³C NMR(126 MHz, DMSO) δ 172.13, 171.40, 169.29, 142.40, 138.78, 134.41,133.23, 130.76, 129.54, 128.47, 127.44, 126.37, 126.12, 125.76, 125.37,124.98, 123.35, 50.76, 49.80, 48.19, 40.72, 40.13, 32.47, 30.27, 28.47,20.96, 18.27.

Example 29 Preparation of (R)-benzyl3-((tert-butoxycarbonyl)amino)-5-(((S)-4-((naphthalen-1-ylmethyl)amino)-4-oxobutan-2-yl)amino)-5-oxopentanoate(PKS2281)

The title compound was synthesized by following the general protocol forHATU mediated coupling of N-Boc-L-beta-glutamic acid 5-benzyl ester (47mg, 0.14 mmol) and H-homo-β-Ala-CH₂-naphth TFA salt (50 mg, 0.14 mmol).The reaction mixture was purified by HPLC to give product (73 mg, 93%)as a white solid. ¹H NMR (500 MHz, DMSO-d₆) δ 8.39 (t, J=5.5 Hz, 1H),8.06-8.04 (m, 1H), 7.95-7.93 (m, 1H), 7.84 (d, J=7.9 Hz, 1H), 7.81 (d,J=8.0 Hz, 1H), 7.56-7.51 (m, 2H), 7.48-7.42 (m, 2H), 7.37-7.30 (m, 5H),6.79 (d, J=8.5 Hz, 1H), 5.05 (s, 2H), 4.76-4.68 (m, 2H), 4.16-4.09 (m,2H), 2.55-2.43 (m, 2H), 2.36 (dd, J=14.0, 5.7 Hz, 1H), 2.26-2.17 (m,3H), 1.35 (s, 9H), 1.02 (d, J=6.0 Hz, 3H).

Example 30 Preparation of(R)-3-((tert-butoxycarbonyl)amino)-5-(((S)-4-((naphthalen-1-ylmethyl)amino)-4-oxobutan-2-yl)amino)-5-oxopentanoicacid (PKS2285)

The title compound was synthesized by following the O-debenzylationprotocol of PKS2281 (73 mg, 0.13 mmol). Yield 61.0 mg (quant.). ¹H NMR(500 MHz, DMSO-d₆) δ 12.12 (s, 1H), 8.39 (t, J=5.6 Hz, 1H), 8.06-8.04(m, 1H), 7.95-7.93 (m, 1H), 7.85 (d, J=8.0 Hz, 1H), 7.79 (d, J=8.0 Hz,1H), 7.56-7.52 (m, 2H), 7.48-7.42 (m, 2H), 6.69 (d, J=8.4 Hz, 1H),4.77-4.68 (m, 2H), 4.15-4.01 (m, 2H), 2.38-2.34 (m, 3H), 2.26-2.16 (m,3H), 1.36 (s, 9H), 1.02 (d, J=6.6 Hz, 3H).

Example 31 Preparation of tert-butyl((R)-1-(tert-butylamino)-5-(((S)-4-((naphthalen-1-ylmethyl)amino)-4-oxobutan-2-yl)amino)-1,5-dioxopentan-3-yl)carbamate(PKS2286)

The title compound was prepared by following the general protocol forEDC mediated coupling of PKS2285 (61.0 mg, 0.13 mmol) andtert-butylamine (20.0 μL, 0.195 mmol). After completion of reaction,mixture was purified by HPLC to give product (13.0 mg, 19%) as a whitesolid. ¹H NMR (500 MHz, DMSO-d₆) δ 8.41 (t, J=5.6 Hz, 1H), 8.06-8.04 (m,1H), 7.95-7.93 (m, 1H), 7.84 (d, J=7.7 Hz, 1H), 7.72 (d, J=8.1 Hz, 1H),7.56-7.51 (m, 2H), 7.48-7.43 (m, 2H), 7.31 (s, 1H), 6.50 (d, J=8.7 Hz,1H), 4.77-4.68 (m, 2H), 4.18-4.10 (m, 1H), 4.06-3.98 (m, 1H), 2.36 (dd,J=13.9, 5.6 Hz, 1H), 2.22-2.14 (m, 5H), 1.36 (s, 9H), 1.22 (s, 9H), 1.02(d, J=6.5 Hz, 3H).

Example 32 Preparation of(R)-3-amino-N¹-(tert-butyl)-N⁵-((S)-4-((naphthalen-1-ylmethyl)amino)-4-oxobutan-2-yl)pentanediamide2,2,2-trifluoroacetate (PKS2288)

The title compound was synthesized by following the general protocol forBoc-deprotection of PKS2286 (13.0 mg, 0.025 mmol). Yield 13.5 mg(quant.). ¹H NMR (500 MHz, Chloroform-d) δ 8.65 (bs, 3H), 8.47 (d, J=7.9Hz, 1H), 7.94 (d, J=8.3 Hz, 1H), 7.86 (d, J=8.1 Hz, 1H), 7.82-7.78 (m,1H), 7.57-7.49 (m, 2H), 7.43-7.40 (m, 2H), 6.52 (t, J=5.3 Hz, 1H), 5.98(s, 1H), 4.81-4.74 (m, 2H), 4.35-4.30 (m, 1H), 3.86 (m, 1H), 2.73-2.68(m, 1H), 2.64-2.58 (m, 1H), 2.46-2.36 (m, 3H), 2.30 (dd, J=14.2, 8.9 Hz,1H), 1.32 (s, 9H), 1.18 (d, J=6.6 Hz, 3H).

Example 33 Preparation of(R)-N¹-(tert-butyl)-3-(4-methylphenylsulfonamido)-N⁵-((S)-4-((naphthalen-1-ylmethyl)amino)-4-oxobutan-2-yl)pentanediamide(PKS2291)

The title compound was prepared by following the general procedure forN-sulfonamide formation of PKS2288 (6.5 mg, 0.012 mmol) with tosylchloride (4.6 mg, 0.024 mmol). The product was isolated as a white solid(5.2 mg, 74%) after HPLC purification. ¹H NMR (500 MHz, DMSO-d₆) δ 8.40(t, J=5.6 Hz, 1H), 8.06-8.04 (m, 1H), 7.96-7.94 (m, 1H), 7.85 (d, J=7.8Hz, 1H), 7.74 (d, J=8.1 Hz, 1H), 7.67-7.66 (m, 2H), 7.55-7.52 (m, 2H),7.47-7.42 (m, 3H), 7.37-7.33 (m, 3H), 4.74 (dd, J=15.2, 5.6 Hz, 1H),4.70 (dd, J=15.2, 5.6 Hz, 1H), 4.11-4.05 (m, 1H), 3.80-3.73 (m, 1H),2.34 (s, 3H), 2.31 (dd, J=13.9, 5.5 Hz, 1H), 2.18-2.08 (m, 5H), 1.18 (s,9H), 0.98 (d, J=6.6 Hz, 3H). ¹³C NMR (126 MHz, DMSO) δ ¹³C NMR (126 MHz,DMSO) δ 169.78, 168.94, 168.36, 142.43, 138.65, 134.60, 133.28, 130.86,129.47, 128.48, 127.56, 126.49, 126.17, 125.80, 125.56, 125.36, 123.52,50.02, 48.52, 42.19, 41.93, 40.54, 40.13, 40.09, 28.42, 20.93, 19.98.

Example 34 Preparation of(R)-N¹-(tert-butyl)-N⁵-((S)-4-((naphthalen-1-ylmethyl)amino)-4-oxobutan-2-yl)-3-(3-phenylpropanamido)pentanediamide(PKS2292)

The title compound was synthesized by following the general protocol forHATU mediated coupling of 3-phenylpropanoic acid (2.0 mg, 0.012) andPKS2288 (6.5 mg, 0.012 mmol). The reaction mixture was purified by HPLCto give product (6.0 mg, 90%) as a white solid. ¹H NMR (500 MHz,DMSO-d₆) δ 8.40 (t, J=5.6 Hz, 1H), 8.06-8.04 (m, 1H), 7.95-7.93 (m, 1H),7.85 (d, J=7.8 Hz, 1H), 7.73 (d, J=8.1 Hz, 1H), 7.64 (d, J=8.3 Hz, 1H),7.56-7.51 (m, 2H), 7.48-7.42 (m, 2H), 7.33 (s, 1H), 7.26-7.23 (m, 2H),7.18-7.14 (m, 3H), 4.76-4.68 (m, 2H), 4.31-4.24 (m, 1H), 4.19-4.10 (m,1H), 2.78 (t, J=7.9 Hz, 2H), 2.38-2.30 (m, 3H), 2.23-2.17 (m, 5H), 1.22(s, 9H), 1.02 (d, J=6.6 Hz, 3H). ¹³C NMR (126 MHz, DMSO) δ 170.46,169.84, 169.42, 168.78, 141.31, 134.61, 133.27, 130.87, 128.47, 128.25,128.09, 127.56, 126.18, 125.82, 125.79, 125.57, 125.37, 123.51, 49.94,44.22, 42.16, 42.04, 40.59, 40.13, 40.06, 37.22, 31.19, 28.48, 20.06.

Example 35 Preparation of PKS3024

The title compound was prepared following the general procedure for EDCmediated coupling of N-Boc-L-β-glutamic acid 5-benzyl ester (168 mg, 0.5mmol) and tert-butylamine (80 μL, 0.75 mmol). After completion ofreaction, water was added to the mixture and extracted twice with ethylacetate. The organic layer was washed with 1N HCl, water, aq. NaHCO3,water, and brine. The organic layer was dried over anhydrous sodiumsulfate and evaporated and dried to give product (196 mg, quant.).Product was used in next step without any further purification. ¹H NMR(500 MHz, Chloroform-d) δ 7.38-7.30 (m, 5H), 5.72 (bs, 1H), 5.59 (bs,1H), 5.12 (s, 2H), 4.24-4.15 (m, 1H), 2.79 (dd, J=16.3, 5.2 Hz, 1H),2.58 (dd, J=15.7, 7.5 Hz, 1H), 2.44-2.34 (m, 2H), 1.41 (s, 9H), 1.30 (s,9H).

Example 36 Preparation of PKS3028

PKS3024 (196 mg, 0.5 mmol) was dissolved in 5 mL 4N HCl (in dioxane) andmixture was stirred at room temperature for 1 hour. After completion ofreaction, dioxane was evaporated and the mixture was diluted with water.The solution was washed with diethyl ether. Aqueous layer was frozen andlyophilized to give product (143 mg, 87%). Complex NMR due to 73:27rotamers. ¹H NMR (500 MHz, DMSO-d₆) δ 8.35-8.26 (m, 2.1H), 8.17 (s,0.9H), 7.87 (s, 1H), 7.48-7.26 (m, 4H), 5.12 (s, 2H), 3.75-3.66 (m,0.73H), 3.66-3.56 (m, 0.27H), 2.83 (dd, J=16.7, 6.6 Hz, 0.73H),2.75-2.65 (m, 1.27H), 2.60-2.52 (m, 2H), 1.24 (d, J=4.5 Hz, 9H).

Example 37 Preparation of PKS3034

The title compound was prepared by following the general procedure forN-sulfonamide formation of PKS3028 (72.3 mg, 0.22 mmol) with tosylchloride (63 mg, 0.33 mmol). The product was isolated by dichloromethaneextraction and purified by column chromatography to give product as awhite solid (50.1 mg, 51%). ¹H NMR (500 MHz, DMSO-d₆) δ 7.73 (d, J=8.2Hz, 1H), 7.65 (d, J=7.8 Hz, 2H), 7.44 (s, 1H), 7.41-7.29 (m, 7H), 4.98(d, J=12.6 Hz, 1H), 4.90 (d, J=12.6 Hz, 1H), 3.88 (dtd, J=13.5, 8.0, 5.3Hz, 1H), 2.46 (dd, J=15.4, 5.4 Hz, 1H), 2.40-2.32 (m, 4H), 2.18 (dd,J=14.6, 8.2 Hz, 1H), 2.12 (dd, J=14.6, 5.4 Hz, 1H), 1.17 (s, 9H).

Example 38 Preparation of PKS3041

The title compound was synthesized by following the general protocol forO-debenzylation of PKS3034 (14.3 mg, 0.032 mmol). Filtrate wasevaporated and dried under vacuum to give product (11.4 mg, quant.). ¹HNMR (500 MHz, DMSO-d₆) δ 7.66 (d, J=7.9 Hz, 2H), 7.42 (s, 1H), 7.35 (d,J=7.9 Hz, 2H), 3.75-3.66 (m, 1H), 2.37 (s, 3H), 2.20-2.11 (m, 4H), 1.18(s, 9H).

Example 39 Preparation of PKS3044

The title compound was prepared following the general procedure for HATUmediated coupling of PKS3041 (11.4 mg, 0.032 mmol) and H-Ala-CH₂-naphthTFA salt (13.1 mg, 0.038 mmol). Purification by HPLC gave product (8.0mg, 44%) as a white solid. ¹H NMR (500 MHz, DMSO-d₆) δ 8.45 (t, J=5.8Hz, 1H), 8.05-8.00 (m, 2H), 7.96-7.92 (m, 1H), 7.83 (d, J=7.9 Hz, 1H),7.67 (d, J=8.0 Hz, 2H), 7.55-7.50 (m, 2H), 7.47-7.40 (m, 3H), 7.39 (s,1H), 7.35 (d, J=8.0 Hz, 2H), 4.74 (dd, J=15.5, 5.7 Hz, 1H), 4.70 (dd,J=15.5, 5.6 Hz, 1H), 4.28-4.18 (m, 1H), 3.86-3.76 (m, 1H), 2.37 (s, 3H),2.22 (d, J=6.8 Hz, 2H), 2.15-2.05 (m, 2H), 1.22-1.15 (m, 12H). ¹³C NMR(126 MHz, DMSO) δ 172.35, 169.18, 168.95, 142.42, 138.82, 134.39,133.23, 130.76, 129.48, 128.47, 127.44, 126.44, 126.13, 125.76, 125.38,125.01, 123.36, 50.08, 48.60, 48.35, 40.75, 40.15, 40.10, 28.40, 20.96,18.09.

Example 40 Preparation of PKS3035

Title compound was synthesized by following the general protocol forHATU mediated coupling of 3-phenylpropanoic acid (40 mg, 0.264 mmol)with PKS3028 (72.3 mg, 0.22 mmol). After completion of reaction, waterwas added to reaction mixture to give white precipitate. Precipitate wasfiltered, washed with water, and dried in air to give product (50 mg,45%) as a white solid. ¹H NMR (500 MHz, DMSO-d₆) δ 7.82 (d, J=8.1 Hz,1H), 7.40 (s, 1H), 7.38-7.28 (m, 5H), 7.28-7.22 (m, 2H), 7.20-7.13 (m,3H), 5.05 (s, 2H), 4.43-4.30 (m, 1H), 2.82-2.68 (m, 2H), 2.55 (dd,J=15.3, 5.6 Hz, 1H), 2.41-2.08 (m, 5H), 1.22 (s, 9H).

Example 41 Preparation of PKS3038

Title compound was prepared following the general protocol forO-debenzylation of PKS3035 (45 mg, 0.106 mmol). The product was isolatedas a white solid (35 mg, quant.). ¹H NMR (500 MHz, DMSO-d₆) δ 7.87 (d,J=8.0 Hz, 1H), 7.44 (s, 1H), 7.29-7.22 (m, 2H), 7.21-7.12 (m, 3H),4.30-4.20 (m, 1H), 2.77 (t, J=7.9 Hz, 2H), 2.37-2.13 (m, 6H), 1.22 (s,9H).

Example 42 Preparation of PKS3039

The title compound was prepared following the general procedure for HATUmediated coupling of PKS3038 (35.0 mg, 0.032 mmol) and H-Ala-CH₂-naphthTFA salt (40.0 mg, 0.116 mmol). Purification by HPLC gave product (17.6mg, 31%) as a white solid. ¹H NMR (500 MHz, DMSO-d₆) δ 8.45 (t, J=5.8Hz, 1H), 8.08-8.01 (m, 2H), 7.97-7.90 (m, 1H), 7.83 (dd, J=7.2, 2.4 Hz,1H), 7.69 (d, J=8.4 Hz, 1H), 7.57-7.49 (m, 2H), 7.49-7.41 (m, 2H), 7.35(s, 1H), 7.27-7.20 (m, 2H), 7.19-7.11 (m, 3H), 4.79-4.69 (m, 2H),4.38-4.27 (m, 2H), 2.75 (t, J=8.0 Hz, 2H), 2.35-2.27 (m, 4H), 2.20 (d,J=6.8 Hz, 2H), 1.24-1.19 (m, 12H). ¹³C NMR (126 MHz, DMSO) δ 172.44,170.56, 169.71, 169.40, 141.29, 134.44, 133.23, 130.77, 128.47, 128.26,128.08, 127.43, 126.14, 125.83, 125.76, 125.38, 124.99, 123.36, 49.99,48.36, 44.32, 40.82, 40.19, 40.01, 37.29, 31.15, 28.46, 18.22.

Example 43 Preparation of PKS3006

Lawesson reagent (61 mg, 0.15 mmol) was suspended in 1 mL THF and asolution of Boc-Ala-CH₂-naphth (98.5 mg, 0.3 mmol) in 2 mL THF was addedat room temperature. The reaction mixture was stirred overnight.Reaction was not complete. Additional 30 mg of Lawesson reaction wasadded and the mixture was stirred additional 18 hours. THF wasevaporated and crude was purified by column chromatography to giveproduct (100 mg, 97%). Complex NMR due to rotamers. ¹H NMR (500 MHz,Chloroform-d) δ 8.31 (s, 1H), 7.96-7.78 (m, 3H), 7.58-7.39 (m, 4H),5.37-5.26 (m, 1H), 5.26-5.10 (m, 2H), 4.56-4.36 (m, 1H), 1.48-1.41 (m,3H), 1.33-1.21 (m, 9H).

Example 44 Preparation of PKS3019

To a stirred solution of PKS3006 (40 mg, 0.116 mmol) in 2 mL ethylacetate, SnCl₄ (1M in heptane, 0.23 mL, 0.230 mmol) was added at roomtemperature. Reaction mixture was stirred at room temperature overnight.Solvent was evaporated and crude was triturated with diethyl ether. Thesolid was filtered and dried to give product as a white solid. The solidwas dissolved in DMF and used in next step. ¹H NMR (500 MHz, DMSO-d₆) δ8.01-7.91 (m, 3H), 7.63-7.46 (m, 5H), 5.25 (d, J=15.2 Hz, 1H), 5.16 (d,J=15.2 Hz, 1H), 4.28-4.17 (m, 1H), 1.40 (d, J=6.6 Hz, 3H).

Example 45 Preparation of PKS3020

The title compound was synthesized by following the general protocol forHATU mediated coupling of PhCH₂CH₂C(O)-Glu(CONHtBu)-OH (18.4 mg, 0.055)and PKS3019 (6.5 mg, 0.05 mmol). The reaction mixture was purified byHPLC to give product (17.3 mg, 62%) as a white solid. ¹H NMR (500 MHz,DMSO-d₆) δ 10.50 (t, J=5.2 Hz, 1H), 8.12 (d, J=7.5 Hz, 1H), 8.09 (d,J=7.7 Hz, 1H), 8.00-7.93 (m, 2H), 7.89 (dd, J=6.5, 3.0 Hz, 1H),7.61-7.51 (m, 2H), 7.51-7.44 (m, 2H), 7.31-7.21 (m, 3H), 7.21-7.13 (m,3H), 5.23-5.18 (m, 2H), 4.78-4.68 (m, 1H), 4.25-4.17 (m, 1H), 2.80 (t,J=8.0 Hz, 2H), 2.49-2.38 (m, 2H), 2.05 (t, J=8.0 Hz, 2H), 1.96-1.85 (m,1H), 1.73-1.62 (m, 1H), 1.32 (d, J=6.8 Hz, 3H), 1.23 (s, 9H). ¹³C NMR(126 MHz, DMSO) δ 204.98, 171.62, 171.16, 170.69, 141.29, 133.29,132.10, 131.01, 128.55, 128.26, 128.13, 127.98, 126.42, 125.99, 125.93,125.83, 125.40, 123.39, 53.87, 52.35, 49.85, 46.77, 36.81, 32.68, 31.06,28.53, 27.77, 21.40.

Example 46 Preparation of PKS3021

The title compound was synthesized by following the general protocol forHATU mediated coupling of Ts-Glu(CONHtBu)-OH (19.6 mg, 0.055) andPKS3019 (6.5 mg, 0.05 mmol). The reaction mixture was purified by HPLCto give product (15.4 mg, 53%) as a white solid. ¹H NMR (500 MHz,DMSO-d₆) δ 10.44 (t, J=5.2 Hz, 1H), 8.18 (d, J=7.4 Hz, 1H), 7.99-7.94(m, 1H), 7.94-7.90 (m, 1H), 7.90-7.83 (m, 2H), 7.63 (d, J=8.3 Hz, 2H),7.58-7.50 (m, 2H), 7.50-7.40 (m, 2H), 7.34-7.29 (m, 3H), 5.20-5.14 (m,2H), 4.51-4.42 (m, 1H), 3.76-3.67 (m, 1H), 2.35 (s, 3H), 2.10-1.95 (m,2H), 1.80-1.70 (m, 1H), 1.64-1.53 (m, 1H), 1.21 (s, 9H), 1.17 (d, J=6.7Hz, 3H). ¹³C NMR (126 MHz, DMSO) δ 204.82, 171.07, 169.63, 142.42,137.96, 133.27, 132.07, 130.98, 129.24, 128.53, 127.95, 126.67, 126.40,125.91, 125.37, 123.37, 55.72, 53.85, 49.86, 46.67, 32.33, 28.77, 28.48,20.99, 20.96.

Example 47 Preparation of PKS3049

The title compound was synthesized following the general protocol forHATU mediated coupling of 3-indoleglyoxylic acid (189 mg, 1.0 mmol) andH-Asp(CONHtBu)-OH TFA salt (432 mg, 1.1 mmol). The compound was isolatedby ethyl acetate extraction and purified by column chromatography togive product (270 mg, 60%) as an off-white solid. ¹H NMR (500 MHz,Chloroform-d) δ 10.14 (s, 1H), 9.13 (d, J=3.3 Hz, 1H), 8.42-8.34 (m,2H), 7.47 (dd, J=6.6, 2.3 Hz, 1H), 7.34-7.23 (m, 7H), 5.54 (s, 1H), 5.23(d, J=12.4 Hz, 1H), 5.19-5.12 (m, 1H), 5.11-5.03 (m, 1H), 2.80 (dd,J=15.2, 5.8 Hz, 1H), 2.74 (dd, J=15.2, 5.3 Hz, 1H), 1.29 (s, 9H). ¹³CNMR (126 MHz, CDCl₃) δ 179.61, 170.76, 168.78, 162.60, 139.33, 136.24,135.30, 128.72, 128.56, 128.41, 126.86, 124.09, 123.31, 122.44, 113.14,112.12, 67.73, 51.99, 49.58, 39.23, 28.78.

Example 48 Preparation of PKS3052

The title compound was synthesized following the general protocol forO-debenzylation of PKS3049 (265 mg, 0.59 mmol). Isolated crude waspurified by HPLC to give product (112 mg, 53%) as an off-white solid. ¹HNMR (500 MHz, DMSO-d₆) δ 12.81 (s, 1H), 12.27 (d, J=3.3 Hz, 1H),8.82-8.75 (m, 2H), 8.26-8.20 (m, 1H), 7.57 (s, 1H), 7.56-7.52 (m, 1H),7.32-7.23 (m, 2H), 4.69-4.60 (m, 1H), 2.67 (dd, J=15.1, 7.2 Hz, 1H),2.59 (dd, J=15.1, 5.0 Hz, 1H), 1.22 (s, 9H). ¹³C NMR (126 MHz, DMSO) δ181.25, 172.26, 168.78, 162.78, 138.64, 136.25, 126.14, 123.53, 122.66,121.28, 112.60, 112.11, 50.16, 48.97, 37.16, 28.44.

Example 49 Preparation of PKS3054

The title compound was synthesized by following the general protocol forHATU mediated coupling of 3-Ind-Glyoxylolyl-Asp-(CONHtBu)-OH (7.2 mg,0.02) and H-β-homo-Ala-CH₂-naphth TFA salt (7.8 mg, 0.022 mmol). Thereaction mixture was purified by HPLC to give product (4.0 mg, 34%) as awhite solid. ¹H NMR (500 MHz, DMSO-d₆) δ 12.24 (d, J=3.6 Hz, 1H), 8.81(d, J=3.1 Hz, 1H), 8.70 (d, J=8.4 Hz, 1H), 8.41 (t, J=5.7 Hz, 1H),8.28-8.21 (m, 1H), 8.02 (dd, J=6.3, 3.4 Hz, 1H), 7.98 (d, J=8.0 Hz, 1H),7.93 (dd, J=6.2, 3.4 Hz, 1H), 7.83 (d, J=8.0 Hz, 1H), 7.58-7.48 (m, 4H),7.48-7.37 (m, 2H), 7.31-7.23 (m, 2H), 4.74 (dd, J=15.2, 5.7 Hz, 1H),4.67 (dd, J=15.2, 5.5 Hz, 1H), 4.62-4.54 (m, 1H), 4.20-4.09 (m, 1H),2.59 (dd, J=14.5, 8.4 Hz, 1H), 2.43 (dd, J=14.5, 4.7 Hz, 1H), 2.36 (dd,J=14.0, 5.8 Hz, 1H), 2.26 (dd, J=14.0, 7.7 Hz, 1H), 1.20 (s, 9H), 1.06(d, J=6.6 Hz, 3H).

Example 50 Preparation of PKS3056

Boc-Ala-OSu (600 mg, 2.10 mmol) was dissolved in 10 mL dichloromethaneand the solution was cooled to 0° C. Aniline and triethylamine wereadded to the solution at 0° C. Reaction mixture was allowed to warm toroom temperature slowly (over 1 hour) and stirred overnight. Water wasadded to the reaction mixture and extracted twice with ethyl acetate.Combined organic layer was washed with 1N HCl and brine, dried overanhydrous sodium sulfate and evaporated to give product 468 mg (84%).Product was used in next step without further purification. ¹H NMR (500MHz, Chloroform-d) δ 8.42 (s, 1H), 7.55-7.48 (m, 2H), 7.31 (td, J=7.5,4.9 Hz, 2H), 7.09 (t, J=7.4 Hz, 1H), 5.14-4.99 (m, 1H), 4.39-4.24 (m,1H), 1.46 (s, 9H), 1.43 (d, J=7.0 Hz, 3H).

Example 51 Preparation of PKS3057

The title compound was synthesized by following the general protocol forBoc-deprotection of Boc-Ala-NHPh (468 mg, 1.77 mmol). The crude obtainedwas suspended in diethyl ether. An off-white solid appeared. Diethylether was decanted and solid was dried to give product (411 mg, 84%).Product was used in next step without further purification. ¹H NMR (500MHz, DMSO-d₆) δ 10.49 (s, 1H), 8.25 (s, 3H), 7.63-7.57 (m, 2H),7.39-7.32 (m, 2H), 7.15-7.08 (m, 1H), 4.02 (d, J=7.2 Hz, 1H), 1.46 (d,J=7.0 Hz, 3H).

Example 52 Preparation of PKS3061

The title compound was synthesized by following the general protocol forHATU mediated coupling of N-Boc-L-β-glutamic acid 5-benzyl ester (84 mg,0.25 mmol) and PKS3057 (83 mg, 0.3 mmol). After completion of reaction,water was added to reaction mixture to give white precipitate.Precipitate was filtered, washed with water, and dried in air to giveproduct (115 mg, 95%) as a white solid. ¹H NMR (500 MHz, DMSO-d₆) δ 9.94(s, 1H), 8.19 (t, J=6.2 Hz, 1H), 7.59 (d, J=8.0 Hz, 2H), 7.44-7.21 (m,7H), 7.04 (t, J=7.4 Hz, 1H), 6.89-6.75 (m, 1H), 5.05 (s, 2H), 4.46-4.32(m, 1H), 4.23-4.06 (m, 1H), 2.62-2.44 (m, 2H), 2.43-2.27 (m, 2H), 1.35(s, 9H), 1.27 (d, J=7.1 Hz, 3H).

Example 53 Preparation of PKS3063

The title compound was synthesized by following the O-debenzylationprotocol of PKS3061 (115 mg, 0.24 mmol) to give product (94.4 mg,quant.) as a white solid. Complex NMR due to 0.28:0.72 ratio ofrotamers. ¹H NMR (500 MHz, DMSO-d₆) δ 12.15 (s, 1H), 10.01 (s, 0.28H),9.95 (s, 0.82H), 8.18 (d, J=7.1 Hz, 1H), 7.62-7.58 (m, 2H), 7.31-7.26(m, 2H), 7.04 (t, J=7.4 Hz, 1H), 6.75 (d, J=8.5 Hz, 0.28H), 6.71 (d,J=8.4 Hz, 0.82H), 4.49-4.31 (m, 1H), 4.16-3.97 (m, 1H), 2.42-2.26 (m,4H), 1.39-1.32 (m, 9H), 1.28 (d, J=7.1 Hz, 3H).

Example 54 Preparation of PKS3065

The title compound was synthesized following the general protocol forEDC mediated coupling of PKS3063 (94.4 mg, 0.24 mmol) andtert-butylamine (38 mL, 0.36 mmol). After completion of reaction waterwas added to give a white precipitate. Precipitate was filtered, washedwith water, and dried in air to give product (85 mg, 79%) as anoff-white solid. ¹H NMR (500 MHz, DMSO-d₆) δ 9.96 (s, 0.28H), 9.93 (s,0.82H), 8.11 (d, J=7.3 Hz, 1H), 7.60 (d, J=8.0 Hz, 2H), 7.35-7.26 (m,3H), 7.04 (t, J=7.4 Hz, 1H), 6.59-6.51 (m, 1H), 4.46-4.36 (m, 1H),4.09-4.00 (m, 1H), 2.37-2.24 (m, 2H), 2.24-2.13 (m, 2H), 1.35 and 1.34(s, 9H), 1.31-1.25 (m, 3H), 1.23 (s, 9H).

Example 55 Preparation of PKS3069

The title compound was synthesized following the general protocol forBoc-deprotection to give 87 mg product (quant.). ¹H NMR (500 MHz,DMSO-d₆) δ 10.05 (s, 1H), 8.57-8.51 (m, 1H), 7.85 (d, J=6.3 Hz, 3H),7.80 (s, 0.76H), 7.79 (s, 0.24H), 7.59 (d, J=8.0 Hz, 2H), 7.34-7.27 (m,2H), 7.05 (t, J=7.4 Hz, 1H), 4.49-4.40 (m, 1H), 3.68-3.60 (m, 1H),2.54-2.46 (m, 2H), 2.45-2.32 (m, 2H), 1.34-1.29 (m, 3H), 1.26 (s, 9H).

Example 56 Preparation of PKS3076

The title compound was synthesized following the general protocol forHATU mediated coupling of 3-phenylpropanoic acid (8.3 mg, 0.055 mmol)and PKS3076 (23.1 mg, 0.05 mmol). Purification by HPLC gave the product(20.0 mg, 83%) as a white solid. Complex NMR due to 0.28:0.72 ratio ofrotamers. ¹H NMR (500 MHz, DMSO-d₆) δ 9.97 (s, 0.28H), 9.94 (s, 0.72H),8.15-8.09 (m, 1H), 7.66 (t, J=7.8 Hz, 1H), 7.62-7.57 (m, 2H), 7.36-7.29(m, 1H), 7.29-7.21 (m, 4H), 7.21-7.12 (m, 3H), 7.07-7.00 (m, 1H),4.46-4.38 (m, 1H), 4.34-4.26 (m, 1H), 2.82-2.72 (m, 2H), 2.40-2.26 (m,4H), 2.24-2.18 (m, 2H), 1.30-1.25 (m, 3H), 1.22 (s, 9H). ¹³C NMR (126MHz, DMSO) δ 171.33, 170.49, 169.79, 169.39, 141.32, 138.94, 128.63,128.23, 128.08, 125.78, 123.26, 119.24, 49.97, 48.97, 44.19, 40.78,39.70, 37.21, 31.14, 28.47, 18.17.

Example 57 Preparation of PKS3077

The title compound was synthesized following the general protocol forHATU mediated coupling of 3-indoleglyoxylic acid (10.4 mg, 0.055 mmol)and PKS3076 (23.1 mg, 0.05 mmol). Purification by HPLC gave the product(15.0 mg, 58%) as an off-white solid. Complex NMR due to 0.27:0.73 ratioof rotamers. ¹H NMR (500 MHz, DMSO-d₆) δ 12.21 (s, 1H), 9.98 (s, 0.27H),9.93 (s, 0.73H), 8.81-8.77 (m, 1H), 8.64 (d, J=8.9 Hz, 1H), 8.28-8.19(m, 2H), 7.63-7.45 (m, 4H), 7.32-7.19 (m, 4H), 7.07-6.97 (m, 1H),4.55-4.46 (m, 1H), 4.45-4.37 (m, 1H), 2.54-2.49 (m, 1H), 2.47-2.34 (m,2H), 2.32-2.25 (m, 1H), 1.29 (d, J=7.1 Hz, 2.3H), 1.24-1.22 (m, 9.7H).¹³C NMR (126 MHz, DMSO) δ 181.66, 171.23, 169.77, 169.36, 162.07,138.85, 138.56, 136.18, 128.58, 126.25, 123.38, 123.23, 122.52, 121.26,119.28, 112.51, 112.10, 50.05, 49.02, 44.19, 40.17, 39.19, 28.43, 18.10.

Example 58 Preparation of PKS3071

The title compound was synthesized by following the general protocol forHATU mediated coupling of Boc-L-β-homoalanine (50.8 mg, 0.25 mmol) and1-naphthylamine hydrochloride (50 mg, 0.275 mmol). After completion ofreaction water was added to give a white precipitate. Precipitate wasfiltered, washed with water, and dried in air to give product (62 mg,76%) as a white solid. ¹H NMR (500 MHz, DMSO-d₆) δ 9.87 (s, 1H),8.09-8.02 (m, 1H), 7.93 (dd, J=6.2, 3.3 Hz, 1H), 7.76 (d, J=8.2 Hz, 1H),7.65 (d, J=7.3 Hz, 1H), 7.57-7.51 (m, 2H), 7.51-7.44 (m, 1H), 6.86 (d,J=8.3 Hz, 1H), 4.06-3.91 (m, 1H), 2.63 (dd, J=13.9, 6.7 Hz, 1H),2.57-2.51 (m, 1H), 1.38 (s, 9H), 1.16 (d, J=6.5 Hz, 3H).

Example 59 Preparation of PKS3073

The title compound was prepared by following the general protocol forBoc-deprotection of PKS3071 (57 mg, 0.174 mmol). The crude wastriturated with diethyl ether and filtered. The solid was dried to giveproduct (59 mg, quant.) as a brown solid. ¹H NMR (500 MHz, DMSO-d₆) δ10.22 (s, 1H), 8.10-8.05 (m, 1H), 8.03-7.92 (m, 4H), 7.79 (d, J=8.2 Hz,1H), 7.69 (d, J=7.4 Hz, 1H), 7.60-7.47 (m, 3H), 3.74-3.58 (m, 1H), 2.85(d, J=6.6 Hz, 2H), 1.31 (d, J=6.6 Hz, 3H).

Example 60 Preparation of PKS21012

The title compound was synthesized by following the general procedurefor the HATU mediated coupling of 3-phenylpropanoic acid (1.68 g, 11.17mmol) with PKS3047 (3.98 g, 10.15 mmol). After completion of reaction,water was added. A white precipitate was formed. The precipitate wasfiltered and washed with water. Precipitate was dried in air to giveproduct (3.92 g, 94%) as a white solid. ¹H NMR (500 MHz, Chloroform-d) δ7.39-7.29 (m, 5H), 7.29-7.23 (m, 2H), 7.21-7.14 (m, 3H), 6.88 (d, J=8.0Hz, 1H), 5.32 (s, 1H), 5.20 (d, J=12.3 Hz, 1H), 5.14 (d, J=12.3 Hz, 1H),4.84-4.77 (m, 1H), 2.95 (t, J=7.9 Hz, 2H), 2.81 (dd, J=15.7, 4.4 Hz,1H), 2.61-2.47 (m, 3H), 1.28 (s, 9H).

Example 61 Preparation of PKS21013

The title compound was synthesized by following the procedure forO-debenzylation of PKS21012 (1.44 g, 3.50 mmol). Product (1.11 g, 99%)was isolated as a white solid. ¹H NMR (500 MHz, DMSO-d₆) δ 12.54 (s,1H), 8.04 (d, J=8.0 Hz, 1H), 7.45 (s, 1H), 7.29-7.23 (m, 2H), 7.23-7.13(m, 3H), 4.52-4.44 (m, 1H), 2.83-2.76 (m, 2H), 2.49-2.44 (m, 1H),2.44-2.34 (m, 3H), 1.22 (s, 9H).

Example 62 Preparation of PKS3081

The title compound was synthesized following the general procedure forHATU mediated coupling of PKS21013 (16.0 mg, 0.05 mmol) and PKS3073(18.8 mg, 0.055 mmol). Purification by HPLC gave the product (21.0 mg,79%) as a white solid. ¹H NMR (500 MHz, DMSO-d₆) δ 9.95 (s, 1H), 8.05(d, J=8.0 Hz, 1H), 8.00 (d, J=8.2 Hz, 1H), 7.96-7.89 (m, 2H), 7.76 (d,J=8.2 Hz, 1H), 7.64 (d, J=7.4 Hz, 1H), 7.58-7.50 (m, 2H), 7.48 (t, J=7.8Hz, 1H), 7.32 (s, 1H), 7.28-7.21 (m, 2H), 7.20-7.12 (m, 3H), 4.60-4.47(m, 1H), 4.32-4.14 (m, 1H), 2.82-2.75 (m, 2H), 2.65 (dd, J=14.2, 5.9 Hz,1H), 2.58 (dd, J=14.2, 7.2 Hz, 1H), 2.45-2.35 (m, 3H), 2.31 (dd, J=14.7,8.6 Hz, 1H), 1.21 (s, 9H), 1.17 (d, J=6.5 Hz, 3H). ¹³C NMR (126 MHz,DMSO) δ 171.15, 170.16, 169.82, 168.77, 141.30, 133.68, 133.44, 128.27,128.12, 128.07, 127.92, 125.97, 125.83, 125.82, 125.50, 125.32, 122.79,122.06, 50.11, 50.05, 42.54, 42.29, 38.80, 36.90, 31.00, 28.45, 20.19.

Example 63 Preparation of PKS21204

Boc-β-Ala-OSu (286.28 mg, 1.00 mmol) was dissolved in dichloromethane(5.00 mL) and 1-naphthylmethanamine (157.21 mg, 1.00 mmol) was added.The solution was cooled to 0° C. and triethylamine (101.19 mg, 1.00mmol) was added. The reaction mixture was allowed to warm to roomtemperature and stirred at room temperature for 2 hours. Aftercompletion of reaction, dichloromethane was evaporated and water wasadded. A white precipitate appeared. Precipitate was filtered, washedwith water and dried in air to give product (320 mg, 97%) as a whitesolid. Product was used in next step without further purification. ¹HNMR (500 MHz, Chloroform-d) δ 7.99 (d, J=8.3 Hz, 1H), 7.88 (d, J=8.0 Hz,1H), 7.85-7.78 (m, 1H), 7.59-7.48 (m, 2H), 7.45-7.40 (m, 2H), 5.90 (bs,1H), 5.14 (bs, 1H), 4.93-4.85 (m, 2H), 3.48-3.37 (m, 2H), 2.45-2.36 (m,2H), 1.39 (s, 9H).

Example 64 Preparation of PKS21211

The title compound was synthesized by following the general procedurefor Boc-deprotection of PKS21204 (320 mg, 0.974 mmol). After completionof reaction, excess trifluoroacetic acid and dichloromethane wereevaporated. Crude was dried under vacuum and triturated with diethylether to give a white solid. Solid was filtered and dried in air to giveproduct (320 mg, 96%). Product was used in next step without furtherpurification. ¹H NMR (500 MHz, DMSO-d₆) δ 8.63 (t, J=5.6 Hz, 1H),8.10-8.02 (m, 1H), 7.96 (dd, J=6.9, 2.2 Hz, 1H), 7.87 (dd, J=7.0, 2.4Hz, 1H), 7.71 (bs, 3H), 7.59-7.52 (m, 2H), 7.51-7.44 (m, 2H), 4.76 (d,J=5.6 Hz, 2H), 3.10-2.96 (m, 2H), 2.53 (t, J=6.8 Hz, 2H).

Example 65 Preparation of PKS21220

The title compound was synthesized by following the general procedurefor HATU mediated coupling of PKS21013 (16.0 mg, 50 umol) and PKS21211(19.6 mg, 74%). After completion of reaction (1 h, temperature rose to10° C.), mixture was purified by preparative LCMS to give product (19.6mg, 74%) as a white solid. ¹H NMR (500 MHz, DMSO-d₆) δ 8.43 (t, J=5.7Hz, 1H), 8.04 (d, J=8.1 Hz, 1H), 7.99 (d, J=8.1 Hz, 1H), 7.94 (d, J=7.8Hz, 1H), 7.84 (d, J=8.0 Hz, 1H), 7.79 (t, J=5.8 Hz, 1H), 7.59-7.50 (m,2H), 7.50-7.40 (m, 2H), 7.34 (s, 1H), 7.28-7.22 (m, 2H), 7.21-7.13 (m,3H), 4.73 (d, J=5.6 Hz, 2H), 4.55-4.45 (m, 1H), 3.31-3.21 (m, 2H), 2.80(t, J=8.0 Hz, 2H), 2.45-2.37 (m, 3H), 2.32 (t, J=7.5 Hz, 2H), 2.30-2.24(m, 1H), 1.21 (s, 9H). ¹³C NMR (126 MHz, DMSO) δ 171.17, 170.88, 170.25,168.76, 141.29, 134.52, 133.27, 130.85, 128.49, 128.27, 128.11, 127.52,126.22, 125.84, 125.77, 125.45, 125.41, 123.43, 50.09, 50.02, 40.15,38.69, 36.92, 35.52, 35.09, 31.00, 28.45.

Example 66 Measurement of IC50s of Dipeptidomimetics AgainstChymotryptic Activities of the Human Constitutive Proteasome andImmunoproteasome

Assays were conducted on a SpectraMax Gemini plate-reader from MolecularDevices (Sunnyvale, Calif.). In a black 96-well plate, 100 μL ofpre-warmed assay mixture containing 0.4 nM Hu i-20S or 0.2 nM Hu c-20S,15 μM N-acetyl-Alanine-Asparagine-Tryptophan-7-amino-4-methylcoumarin(Ac-ANW-AMC) (substrate for i-20S) or 25 μMN-succinyl-Leucine-Leucine-Valine-Tyrosine-7-amino-4-methylcoumarin(suc-LLVY-AMC) (substrate for c-20S), 0.02% SDS in buffer (50 mM HEPES,0.5 mM EDTA, pH 7.4) was added to the wells that contained 1 μLinhibitor at 100× indicated concentrations in DMSO. The reactionprogress of each well was recorded by monitoring fluorescence at 460 nm(λ_(ex)=360 nm) for 120 minutes at 37° C. Velocities were derived fromthe initial linear range of the curves and the IC50s were obtained byplotting the percentage of inhibitor versus inhibitor concentrations(FIG. 1).

Example 67 Proteasome Inhibitory Studies of Various N,C CappedDipeptidomimetics

A variety of dipeptidomimetics were synthesized and tested for theirability to inhibit the proteasomes, as described in Example 1. Thesecompounds inhibited the proteasomes reversibly. The IC50s of thesecompounds against human immunoproteasome β5i and constitutive proteasomeβ5c are listed in Table 1. IC50 curves are shown in FIG. 1. These datashow that compounds with substituents at amide side chain of theAsparagine (2nd amino acid from C-terminus) and C-terminal caps allowedthe resulting dipeptides to selectively inhibit the humanimmunoproteasome β5i active site over the human constitutive proteasomeβ5c active site.

TABLE 1 Kinetic parameters and calculated logP of N,C-cappeddipeptidomimetics IC50 (μM) ID Structures Hu i-20S Hu c-20S logP PKS2249

0.18 16.1 0.55 PKS2251

0.045 32.2 2.75 PKS2252

0.0064 8.81 2.72 PKS2253

0.31 17.8 3.03 PKS2260

0.0112 16.55 2.23 PK52272

0.152 >100 1.95 PKS2278

2.26 >100 2.75 PK52279

0.0145 34.7 3.73 PK52290

1.60 13.9 4.19 PK52291

8.68 >100 4.19 PKS2292

>100 >100 2.87 PK52295

0.141 ND 0.69 PKS3020

0.799 7.0 3.9 PK53021

0.671 13.4 3.86 PKS3054

0.0048 0.234 1.47 PKS3065

>100 >100 0.89 PKS3081

13.7 >100 2.68 PKS21220

0.655 51.4 2.59

Although the invention has been described in detail, for the purpose ofillustration, it is understood that such detail is for that purpose andvariations can be made therein by those skilled in the art withoutdeparting from the spirit and scope of the invention which is defined bythe following claims.

1.-7. (canceled)
 8. A method of treating cancer, immunologic disorders,autoimmune disorders, neurodegenerative disorders, or inflammatorydisorders in a subject or for providing immunosuppression fortransplanted organs or tissues in a subject, said method comprising:administering to the subject in need thereof a compound of the Formula(I):

wherein L is —(CR³R^(x))_(p)—; M is —(CR²R^(y))_(r)—; R¹ is selectedfrom the group consisting of monocyclic and bicyclic aryl, biphenyl,monocyclic and bicyclic heteroaryl, monocyclic and bicyclicheterocyclyl, and monocyclic and bicyclic non-aromatic heterocycle,wherein monocyclic and bicyclic aryl, biphenyl, monocyclic and bicyclicheteroaryl, monocyclic and bicyclic heterocyclyl, and monocyclic andbicyclic non-aromatic heterocycle can be optionally substituted from 1to 3 times with a substituent selected independently at each occurrencethereof from the group consisting of halogen, cyano, —CF₃, C₁₋₆ alkyl,and C₁₋₆ alkoxy; R² is independently selected at each occurrence thereoffrom the group consisting of H, D, C₁₋₆ alkyl, —CH₂OC₁₋₆ alkyl, —CH₂Ar,and —CH₂heteroaryl, wherein aryl (Ar) can be optionally substituted from1 to 3 times with a substituent selected independently at eachoccurrence thereof from the group consisting of halogen, cyano, C₁₋₆alkyl, and C₁₋₆ alkoxy; R³ is independently selected at each occurrencethereof from the group consisting of H, D, —CH₂OC₁₋₆ alkyl,—(CH₂)_(m)C(O)NHR⁵, —(CH₂)_(m)C(O)NR⁶R⁷, —(CH₂)_(m)C(O)OH, and—(CH₂)_(m)C(O)OBn_(;) R⁴ is selected from the group consisting of H,—C(O)(CH₂)_(n)Ph, —C(O)CH₂NR⁶R⁷, —SO₂Ar, —SO₂C₁₋₆ alkyl, —SO₂C₃₋₆cycloalkyl, —C(O)(CH₂)_(n)Het, —C(O)C(O)Het, —C(O)C₁₋₆ alkyl, —C(O)OC₁₋₆alkyl, —C(O)CF₃, heteroaryl, and —(CH₂)_(n)NR⁶R⁷, wherein aryl (Ar) andheteroaryl (Het) can be optionally substituted from 1 to 3 times with asubstituent selected independently at each occurrence thereof from thegroup consisting of halogen, cyano, C₁₋₆ alkyl, and C₁₋₆ alkoxy; R⁵ isselected from the group consisting of C₁₋₆ alkyl, C₁₋₆ alkoxy,non-aromatic heterocycle, —NR⁶R⁷, and —CR⁸R⁹; R⁶, R⁷, R⁸, and R⁹ areeach independently selected from the group consisting of H, D, C₁₋₆alkyl, and —(CH₂)_(k)OH; or R⁶ and R⁷ are taken together with thenitrogen to which they are attached to form a morpholine ring; or R⁸ andR⁹ are taken together with the carbon to which they are attached to forman oxetane ring; R^(x) is independently selected at each occurrencethereof from the group consisting of H, D, —CH₂OC₁₋₆ alkyl,—(CH₂)_(m)C(O)NHR⁵, —(CH₂)_(m)C(O)NR⁶R⁷, and —CH₂C(O)R⁵ _(;) R^(y) isindependently selected at each occurrence thereof from the groupconsisting of H, D, C₁₋₆ alkyl, —CH₂OC₁₋₆ alkyl, —CH₂Ar, and—CH₂heteroaryl, wherein aryl (Ar) can be optionally substituted from 1to 3 times with a substituent selected independently at each occurrencethereof from the group consisting of halogen, cyano, C₁₋₆ alkyl, andC₁₋₆ alkoxy; X is —(CH₂)_(q)—, —O—, or —(CD₂)_(q)—; Y is O or S; k is 1,2, or 3; m is 0, 1, 2, 3, 4, or 5; n is 0, 1, 2, or 3; p is 1 or 2; q is0, 1, or 2; r is 1 or 2; and s is 0 or 1; with a proviso that when s is0, then r is 2; and when s is 1, then r+p≥3.
 9. The method of claim 8,wherein the compound of Formula (I) has the formula:


10. The method of claim 8, wherein R¹ is selected from the groupconsisting of

and R¹¹ is selected from the group consisting of halogen, cyano, —CF₃,C₁₋₆ alkyl, and C₁₋₆ alkoxy.
 11. The method of claim 8, wherein R² isselected from the group consisting of H, Me, —CH₂(Me)₂, —CH₂OMe,


12. The method of claim 8, wherein R³ is selected from the groupconsisting of —CH₂OMe, —CH₂C(O)OH, —CH₂C(O)OBn, —(CH₂)₂C(O)OBn,—(CH₂)₂C(O)OH,


13. The method of claim 8, wherein R⁴ is selected from the groupconsisting of H, trifluoroacetyl,

l is 0, 1, 2, 3, or 4; m is 0, 1, 2, 3, 4, or 5; n is 0, 1, 2, or 3; andR is selected from the group consisting of H, halogen, cyano, C₁₋₆alkyl, and C₁₋₆ alkoxy.
 14. The method of claim 8, wherein the compoundof Formula (I) is selected from the group consisting of:


15. The method of claim 8, wherein an autoimmune disorder is treated,said autoimmune disorder being selected from the group consisting ofarthritis, colitis, multiple sclerosis, lupus, systemic sclerosis, andsjögren syndrome.
 16. The method of claim 8, wherein immunosuppressionis provided for transplanted organs or tissues, said immunosuppressionbeing used to prevent transplant rejection and graft-verse-host disease.17. The method of claim 8, wherein an inflammatory disorder is treated,said inflammatory disorder being Crohn's disease and ulcerative colitis.18. The method of claim 8, wherein cancer is treated, said cancer beingselected from the group consisting of neoplastic disorders, hematologicmalignancies, and lymphocytic malignancies. 19.-26. (canceled)
 27. Amethod of treating cancer, immunologic disorders, autoimmune disorders,neurodegenerative disorders, or inflammatory disorders in a subject orfor providing immunosuppression for transplanted organs or tissues in asubject, said method comprising: administering to the subject in needthereof a compound of the Formula (Ia), Formula (Ib), or Formula (Ic):

wherein R¹ is selected from the group consisting of monocyclic andbicyclic aryl, biphenyl, monocyclic and bicyclic heteroaryl, monocyclicand bicyclic heterocyclyl, and monocyclic and bicyclic non-aromaticheterocycle, wherein monocyclic and bicyclic aryl, biphenyl, monocyclicand bicyclic heteroaryl, monocyclic and bicyclic heterocyclyl, andmonocyclic and bicyclic non-aromatic heterocycle can be optionallysubstituted from 1 to 3 times with a substituent selected independentlyat each occurrence thereof from the group consisting of halogen, cyano,—CF₃, C₁₋₆ alkyl, and C₁₋₆ alkoxy; R² is independently selected at eachoccurrence thereof from the group consisting of H, D, C₁₋₆ alkyl,—CH₂OC₁₋₆ alkyl, —CH₂Ar, and —CH₂heteroaryl, wherein aryl (Ar) can beoptionally substituted from 1 to 3 times with a substituent selectedindependently at each occurrence thereof from the group consisting ofhalogen, cyano, C₁₋₆ alkyl, and C₁₋₆ alkoxy; R³ is independentlyselected at each occurrence thereof from the group consisting of H, D,—CH₂OC₁₋₆ alkyl, —(CH₂)_(m)C(O)NHR⁵, and —(CH₂)_(m)C(O)NR⁶R⁷ _(;) R⁴ isselected from the group consisting of —C(O)(CH₂)_(n)Ph, —C(O)CH₂NR⁶R⁷,—SO₂Ar, —SO₂C₁₋₆ alkyl, —SO₂C₃₋₆ cycloalkyl, —C(O)(CH₂)_(n)Het,—C(O)C(O)Het, —C(O)C₁₋₆ alkyl, —C(O)OC₁₋₆ alkyl, —C(O)CF₃, heteroaryl,and —(CH₂)_(n)NR⁶R⁷, wherein aryl (Ar) and heteroaryl (Het) can beoptionally substituted from 1 to 3 times with a substituent selectedindependently at each occurrence thereof from the group consisting ofhalogen, cyano, C₁₋₆ alkyl, and C₁₋₆ alkoxy; R⁵ is selected from thegroup consisting of C₁₋₆ alkyl, C₁₋₆ alkoxy, non-aromatic heterocycle,—NR⁶R⁷, and —CR⁸R⁹; R⁶, R⁷, R⁸, and R⁹ are each independently selectedfrom the group consisting of H, D, C₁₋₆ alkyl, and —(CH₂)_(k)OH; or R⁶and R⁷ are taken together with the nitrogen to which they are attachedto form a morpholine ring; or R⁸ and R⁹ are taken together with thecarbon to which they are attached to form an oxetane ring; R^(x) isindependently selected at each occurrence thereof from the groupconsisting of H, D, —CH₂OC₁₋₆ alkyl, —(CH₂)_(m)C(O)NHR⁵,—(CH₂)_(m)C(O)NR⁶R⁷, and —CH₂C(O)R⁵ _(;) R^(y) is independently selectedat each occurrence thereof from the group consisting of H, D, C₁₋₆alkyl, —CH₂OC₁₋₆ alkyl, —CH₂Ar, and —CH₂heteroaryl, wherein aryl (Ar)can be optionally substituted from 1 to 3 times with a substituentselected independently at each occurrence thereof from the groupconsisting of halogen, cyano, C₁₋₆ alkyl, and C₁₋₆ alkoxy; X is—(CH₂)_(q)—, —O—, or —(CD₂)_(q)—; Y is O or S; k is 1, 2, or 3; m is 0,1, 2, 3, 4, or 5; n is 0, 1, 2, or 3; q is 0, 1, or 2; and s is 0 or 1.28. The method of claim 27, wherein the compound of Formula (Ia),Formula (Ib), or Formula (Ic) has the formula:


29. The method of claim 27, wherein R¹ is selected from the groupconsisting of

and R¹¹ is selected from the group consisting of halogen, cyano, —CF₃,C₁₋₆ alkyl, and C₁₋₆ alkoxy.
 30. The method of claim 27, wherein R² isselected from the group consisting of Me, —CH₂(Me)₂, —CH₂OMe,


31. The method of claim 27, wherein R³ is selected from the groupconsisting of —CH₂OMe,


32. The method of claim 27, wherein R⁴ is selected from the groupconsisting of trifluoroacetyl,

l is 0, 1, 2, 3, or 4; m is 0, 1, 2, 3, 4, or 5; n is 0, 1, 2, or 3; andR is selected from the group consisting of H, halogen, cyano, C₁₋₆alkyl, and C₁₋₆ alkoxy.
 33. The method of claim 27, wherein the compoundof Formula (Ia), Formula (Ib), or Formula (Ic) is selected from thegroup consisting of:


34. The method of claim 27, wherein an autoimmune disorder is treated,said autoimmune disorder being selected from the group consisting ofarthritis, colitis, multiple sclerosis, and lupus.
 35. The method ofclaim 27, wherein immunosuppression is provided for transplanted organsor tissues, said immunosuppression being used to prevent transplantrejection and graft-verse-host disease.
 36. The method of claim 27,wherein an inflammatory disorder is treated, said inflammatory disorderbeing Crohn's disease.
 37. The method of claim 27, wherein cancer istreated, said cancer being selected from the group consisting ofneoplastic disorders, hematologic malignancies, and lymphocyticmalignancies.
 38. (canceled)